Edinburgh/Notebook/Minipreps

From 2008.igem.org

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<html>
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<div id="header">{{Template:Team:Edinburgh/Templates/Header}}</div>
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<head>
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<title>Edinburgh iGem 2008</title>
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<script type="text/javascript">
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//Drop Down Tabs Menu- Author: Dynamic Drive (http://www.dynamicdrive.com)
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//Created: May 16th, 07'
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var tabdropdown={
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'''[[Team:Edinburgh/Notebook|< Back to Notebook]]'''
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* disappeardelay: 200, //set delay in miliseconds before menu disappears onmouseout
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disablemenuclick: false, //when user clicks on a menu item with a drop down menu, disable menu item's link?
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enableiframeshim: 1, //1 or 0, for true or false
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//No need to edit beyond here////////////////////////
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== Minipreps of Confirmed Clones ==
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dropmenuobj: null, ie: document.all, firefox: document.getElementById&&!document.all, previousmenuitem:null,
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currentpageurl: window.location.href.replace("http://"+window.location.hostname, "").replace(/^\//, ""), //get current page url (minus hostname, ie: http://www.dynamicdrive.com/)
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getposOffset:function(what, offsettype){
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* '''M2:''' pSB1A2+''dxs''
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var totaloffset=(offsettype=="left")? what.offsetLeft : what.offsetTop;
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* '''M15:''' pSB1A2+''appY''
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var parentEl=what.offsetParent;
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* '''(?) M36:''' pSB1A2+''crtB'' (sequencing results not recorded, but follow-up did occur)
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while (parentEl!=null){
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* '''(?) M42:''' pSB1A2+''crtI'' (sequencing results not recorded, but follow-up did occur)
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totaloffset=(offsettype=="left")? totaloffset+parentEl.offsetLeft : totaloffset+parentEl.offsetTop;
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* '''M43:''' BABEL2(?)+''glgC'' double mutant.
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parentEl=parentEl.offsetParent;
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* '''M50:''' pSB1A2+rbs+''crtI''
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}
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* '''M63:''' BABEL2+rbs+''crtE''
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return totaloffset;
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* '''M67:''' pSB1A2+rbs+''crtB''
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},
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* '''M72:''' pSB1A2+rbs+''dxs''
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* '''M79:''' BABEL2+''crtE''
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* '''M82:''' pSB1A2+rbs+''appY''
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* '''M109:''' pSB1A2+''crtBI''
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* '''(?) M121:''' pSB1A2+''crtY'' (sequencing results not posted; no follow-up recorded)
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* '''(?) M124''' pSB1A2+rbs+''crtY'' (sequencing results not posted; no follow-up recorded)
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* '''M116:''' pSB1A2+''glgC'' double mutant
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* '''M130:''' pSB1A2+P''cstA''
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* '''M137:''' pSB1A2+''dxs+LIMS1''
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* '''M150:''' pSB1A2+''cex''
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* '''M152:''' pSB1A2+''dxs+crtE''
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** '''M154:''' not sequenced, but similar to M152.
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** '''M155:''' not sequenced, but also similar to M152; ligated to M110 (''crtBI'') in L62.
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* '''M162:''' pSB1A2+''dxs+LIMS1+appY''
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* '''M212:''' pSB1A2+''glgC'' triple mutant (ie. ''glgC16'')
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* '''M222:''' pSB1A2+P''lac''+''dxs+LIMS1+appY''
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* '''M233:''' pSB1A2+''cex'' mutant (PstI site removed)
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** '''M234, 235:''' not sequenced, but similar to M233.
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* '''M237, 238:''' pSB1A2+''cenA''
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* '''M243:''' pSB1A2+rbs+''glgC''
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* '''M245:''' pSB1A2+rbs+''glgC16''
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* '''M254:''' pSB1A2+P''lac''+rbs+''glgC''
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* '''M256:''' pSB1A2+P''lac''+rbs+''glgC16''
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* '''M258:''' pSB1A2+P''cstA''+''lacZ' '' (BBa_J15202)
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* '''M259:''' pSB1A2+P''cstA''+''xylE'' (BBa_J33204)
 +
* '''M281:''' pSB1A2+''bglX'' coding sequence
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* '''M303:''' pSB1A2+rbs+''cex''
 +
* '''M305:''' pSB1A2+rbs+''cenA''
 +
* '''M307:''' pSB1A2+rbs+''bglX''
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showhide:function(obj, e, obj2){ //obj refers to drop down menu, obj2 refers to tab menu item mouse is currently over
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== Complete List of Minipreps ==
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if (this.ie || this.firefox)
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this.dropmenuobj.style.left=this.dropmenuobj.style.top="-500px"
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if (e.type=="click" && obj.visibility==hidden || e.type=="mouseover"){
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if (obj2.parentNode.className.indexOf("default")==-1) //if tab isn't a default selected one
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obj2.parentNode.className="selected"
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obj.visibility="visible"
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}
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else if (e.type=="click")
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obj.visibility="hidden"
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},
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iecompattest:function(){
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return (document.compatMode && document.compatMode!="BackCompat")? document.documentElement : document.body
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},
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clearbrowseredge:function(obj, whichedge){
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var edgeoffset=0
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if (whichedge=="rightedge"){
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var windowedge=this.ie && !window.opera? this.standardbody.scrollLeft+this.standardbody.clientWidth-15 : window.pageXOffset+window.innerWidth-15
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this.dropmenuobj.contentmeasure=this.dropmenuobj.offsetWidth
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if (windowedge-this.dropmenuobj.x < this.dropmenuobj.contentmeasure)  //move menu to the left?
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edgeoffset=this.dropmenuobj.contentmeasure-obj.offsetWidth
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}
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else{
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var topedge=this.ie && !window.opera? this.standardbody.scrollTop : window.pageYOffset
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var windowedge=this.ie && !window.opera? this.standardbody.scrollTop+this.standardbody.clientHeight-15 : window.pageYOffset+window.innerHeight-18
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this.dropmenuobj.contentmeasure=this.dropmenuobj.offsetHeight
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if (windowedge-this.dropmenuobj.y < this.dropmenuobj.contentmeasure){ //move up?
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edgeoffset=this.dropmenuobj.contentmeasure+obj.offsetHeight
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if ((this.dropmenuobj.y-topedge)<this.dropmenuobj.contentmeasure) //up no good either?
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edgeoffset=this.dropmenuobj.y+obj.offsetHeight-topedge
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}
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this.dropmenuobj.firstlink.style.borderTopWidth=(edgeoffset==0)? 0 : "1px" //Add 1px top border to menu if dropping up
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}
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return edgeoffset
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},
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dropit:function(obj, e, dropmenuID){
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if (this.dropmenuobj!=null){ //hide previous menu
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this.dropmenuobj.style.visibility="hidden" //hide menu
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if (this.previousmenuitem!=null && this.previousmenuitem!=obj){
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if (this.previousmenuitem.parentNode.className.indexOf("default")==-1) //If the tab isn't a default selected one
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this.previousmenuitem.parentNode.className=""
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}
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}
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this.clearhidemenu()
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if (this.ie||this.firefox){
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obj.onmouseout=function(){tabdropdown.delayhidemenu(obj)}
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obj.onclick=function(){return !tabdropdown.disablemenuclick} //disable main menu item link onclick?
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this.dropmenuobj=document.getElementById(dropmenuID)
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this.dropmenuobj.onmouseover=function(){tabdropdown.clearhidemenu()}
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this.dropmenuobj.onmouseout=function(e){tabdropdown.dynamichide(e, obj)}
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this.dropmenuobj.onclick=function(){tabdropdown.delayhidemenu(obj)}
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this.showhide(this.dropmenuobj.style, e, obj)
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this.dropmenuobj.x=this.getposOffset(obj, "left")
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this.dropmenuobj.y=this.getposOffset(obj, "top")
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this.dropmenuobj.style.left=this.dropmenuobj.x-this.clearbrowseredge(obj, "rightedge")+"px"
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this.dropmenuobj.style.top=this.dropmenuobj.y-this.clearbrowseredge(obj, "bottomedge")+obj.offsetHeight+1+"px"
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this.previousmenuitem=obj //remember main menu item mouse moved out from (and into current menu item)
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this.positionshim() //call iframe shim function
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-
}
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},
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contains_firefox:function(a, b) {
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while (b.parentNode)
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if ((b = b.parentNode) == a)
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return true;
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return false;
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},
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dynamichide:function(e, obj2){ //obj2 refers to tab menu item mouse is currently over
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var evtobj=window.event? window.event : e
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if (this.ie&&!this.dropmenuobj.contains(evtobj.toElement))
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this.delayhidemenu(obj2)
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else if (this.firefox&&e.currentTarget!= evtobj.relatedTarget&& !this.contains_firefox(evtobj.currentTarget, evtobj.relatedTarget))
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-
this.delayhidemenu(obj2)
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},
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delayhidemenu:function(obj2){
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this.delayhide=setTimeout(function(){tabdropdown.dropmenuobj.style.visibility='hidden'; if (obj2.parentNode.className.indexOf('default')==-1) obj2.parentNode.className=''},this.disappeardelay) //hide menu
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},
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clearhidemenu:function(){
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if (this.delayhide!="undefined")
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clearTimeout(this.delayhide)
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},
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positionshim:function(){ //display iframe shim function
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if (this.enableiframeshim && typeof this.shimobject!="undefined"){
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-
if (this.dropmenuobj.style.visibility=="visible"){
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-
this.shimobject.style.width=this.dropmenuobj.offsetWidth+"px"
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-
this.shimobject.style.height=this.dropmenuobj.offsetHeight+"px"
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this.shimobject.style.left=this.dropmenuobj.style.left
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this.shimobject.style.top=this.dropmenuobj.style.top
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}
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this.shimobject.style.display=(this.dropmenuobj.style.visibility=="visible")? "block" : "none"
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}
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},
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hideshim:function(){
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if (this.enableiframeshim && typeof this.shimobject!="undefined")
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this.shimobject.style.display='none'
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},
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isSelected:function(menuurl){
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var menuurl=menuurl.replace("http://"+menuurl.hostname, "").replace(/^\//, "")
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return (tabdropdown.currentpageurl==menuurl)
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},
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init:function(menuid, dselected){
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this.standardbody=(document.compatMode=="CSS1Compat")? document.documentElement : document.body //create reference to common "body" across doctypes
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var menuitems=document.getElementById(menuid).getElementsByTagName("a")
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for (var i=0; i<menuitems.length; i++){
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if (menuitems[i].getAttribute("rel")){
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var relvalue=menuitems[i].getAttribute("rel")
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document.getElementById(relvalue).firstlink=document.getElementById(relvalue).getElementsByTagName("a")[0]
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menuitems[i].onmouseover=function(e){
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var event=typeof e!="undefined"? e : window.event
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tabdropdown.dropit(this, event, this.getAttribute("rel"))
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-
}
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}
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if (dselected=="auto" && typeof setalready=="undefined" && this.isSelected(menuitems[i].href)){
+
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menuitems[i].parentNode.className+=" selected default"
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-
var setalready=true
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}
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else if (parseInt(dselected)==i)
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menuitems[i].parentNode.className+=" selected default"
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-
}
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}
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-
 
+
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}
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</script>
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<style>
+
-
.ddcolortabs{
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-
padding: 0;
+
-
width: 100%;
+
-
background: transparent;
+
-
voice-family: "\"}\"";
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voice-family: inherit;
+
-
}
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-
 
+
-
.ddcolortabs ul{
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font: normal 11px Arial, Verdana, sans-serif;
+
-
margin:0;
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-
padding:0;
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-
list-style:none;
+
-
}
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-
 
+
-
.ddcolortabs li{
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display:inline;
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-
margin:0 2px 0 0;
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-
padding:0;
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-
text-transform:uppercase;
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-
}
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-
 
+
-
 
+
-
.ddcolortabs a{
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float:left;
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-
color: white;
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-
background: black url(media/color_tabs_left.gif) no-repeat left top;
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-
margin:0 2px 0 0;
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-
padding:0 0 1px 3px;
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-
text-decoration:none;
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-
letter-spacing: 1px;
+
-
}
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-
 
+
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.ddcolortabs a span{
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-
float:left;
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display:block;
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-
background: transparent url(media/color_tabs_right.gif) no-repeat right top;
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-
padding: 4px 8px 2px 7px;
+
-
}
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-
 
+
-
.ddcolortabs a span{
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-
float:none;
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-
}
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-
.ddcolortabs a:hover{
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-
background-color: #591f20;
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-
}
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-
 
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-
.ddcolortabs a:hover span{
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-
background-color: #591f20;
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-
}
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.ddcolortabs .selected a, #ddcolortabs .selected a span{ /*currently selected tab*/
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-
background-color: #591f20;
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-
}
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-
 
+
-
.ddcolortabsline{
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-
clear: both;
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-
padding: 0;
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-
width: 100%;
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-
height: 8px;
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-
line-height: 8px;
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-
background: black;
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-
border-top: 1px solid #fff; /*Remove this to remove border between bar and tabs*/
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-
}
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-
 
+
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/* ######### Style for Drop Down Menu ######### */
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-
 
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.dropmenudiv_a{
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-
position:absolute;
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top: 0;
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border: 1px solid black; /*THEME CHANGE HERE*/
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border-top-width: 8px; /*Top border width. Should match height of .ddcolortabsline above*/
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border-bottom-width: 0;
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-
font:normal 12px Arial;
+
-
line-height:18px;
+
-
z-index:100;
+
-
background-color: white;
+
-
width: 200px;
+
-
visibility: hidden;
+
-
}
+
-
 
+
-
 
+
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.dropmenudiv_a a{
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-
width: auto;
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-
display: block;
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-
text-indent: 5px;
+
-
border-top: 0 solid #678b3f;
+
-
border-bottom: 1px solid #678b3f; /*THEME CHANGE HERE*/
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-
padding: 2px 0;
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text-decoration: none;
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color: black;
+
-
}
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+
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* html .dropmenudiv_a a{ /*IE only hack*/
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-
width: 100%;
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}
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+
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.dropmenudiv_a a:hover{ /*THEME CHANGE HERE*/
+
-
background-color: #8a3c3d;
+
-
color: white;
+
-
}
+
-
</style>
+
-
</head>
+
-
 
+
-
<!-- CSS for Drop Down Tabs Menu #1 -->
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<link rel="stylesheet" type="text/css" href="ddcolortabs.css" />
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<div id="colortab" class="ddcolortabs">
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<ul>
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<li><a href="https://2008.igem.org/Team:Edinburgh" title="Home"><span>Home</span></a></li>
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<li><a href="https://2008.igem.org/Team:Edinburgh/Project" title="Project" rel="dropmenu1_a"><span>The Project</span></a></li>
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<li><a href="https://2008.igem.org/Team:Edinburgh/Team" title="Team" ><span>The Team</span></a></li>
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<li><a href="http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=Edinburgh" title="Project" rel="dropmenu1_a"><span>BioBrick Parts</span></a></li>
+
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<li><a href="https://2008.igem.org/Team:Edinburgh/Modeling" title="Modelling" rel="dropmenu2_a"><span>Modelling</span></a></li>
+
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<li><a href="https://2008.igem.org/Team:Edinburgh/Notebook" title="Notebook"><span>Notebook</span></a></li>
+
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</ul>
+
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</div>
+
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<div class="ddcolortabsline">&nbsp;</div>
+
-
<div id="dropmenu1_a" class="dropmenudiv_a">
+
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<a href="https://2008.igem.org/Team:Edinburgh/Team">Overview</a>
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<a href="https://2008.igem.org/Team:Edinburgh/Team">Step1</a>
+
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<a href="https://2008.igem.org/Team:Edinburgh/Team">Step2</a>
+
-
</div>
+
-
<script type="text/javascript">
+
-
//SYNTAX: tabdropdown.init("menu_id", [integer OR "auto"])
+
-
tabdropdown.init("colortab", 3)
+
-
</script>
+
-
</html>
+
'''M1 to M6:''' pSB1A2+''dxs'' transformants (plate 11). M2, M3, M4 and M6 all looked correct on gel 2. (30.06.08) - M2 was sequenced to confirm identity and was maxiprepped as '''X2'''.<br />
'''M1 to M6:''' pSB1A2+''dxs'' transformants (plate 11). M2, M3, M4 and M6 all looked correct on gel 2. (30.06.08) - M2 was sequenced to confirm identity and was maxiprepped as '''X2'''.<br />
'''M7:''' BABEL1+''glgC'' transformant (plate 12). Did not look right on gel. (30.06.08)<br />
'''M7:''' BABEL1+''glgC'' transformant (plate 12). Did not look right on gel. (30.06.08)<br />
'''M8 to M12:''' BABEL2+''glgC'' transformants (plate 13). M10 and M11 both looked good on a gel, but sequencing showed that both had the insert in the reverse orientation. (30.06.08)<br />
'''M8 to M12:''' BABEL2+''glgC'' transformants (plate 13). M10 and M11 both looked good on a gel, but sequencing showed that both had the insert in the reverse orientation. (30.06.08)<br />
-
'''M13 to M18:''' pSB1A2+''appY'' transformants. Gel 10 showed that M15 to M18, and possibly M14, all seemed to have inserts the right size.<br />
+
'''M13 to M18:''' pSB1A2+''appY'' transformants. Gel 10 showed that M15 to M18, and possibly M14, all seemed to have inserts the right size. (09.07.08)<br />
-
'''M19 to M24:''' Babel2+''glgC'' clones after mutation of EcoRI site 2. EcoRI digests on *Gel 11* suggest that *M19, M21 and M22* may have lost the EcoRI site.<br />
+
'''M19 to M24:''' BABEL2+''glgC'' clones after mutation of EcoRI site 2. EcoRI digests on *Gel 11* suggest that *M19, M21 and M22* may have lost the EcoRI site. (10.07.08)<br />
-
'''M25 to M30:''' possible rbs+''dxs'' transformants.<br />
+
'''M25 to M30:''' possible rbs+''dxs'' transformants from plate 34. (13.07.08)<br />
-
'''M31 to M36:''' ''crtB'' clones.<br />
+
'''M31 to M36:''' pSB1A2+''crtB'' clones from plate 35. (14.07.08)<br />
-
'''M37 to M42:''' ''crtI'' clones.<br />
+
'''M37 to M42:''' pSB1A2+''crtI'' clones from plate 36. (14.07.08)<br />
-
'''M43 to M47:''' possible ''glgC'' double mutants (both EcoRI sites gone)<br />
+
'''M43 to M47:''' possible BABEL2+''glgC'' double mutants (both EcoRI sites gone). (19.07.08)<br />
-
'''M48:''' a possible rbs+''dxs'' clone.<br />
+
'''M48:''' a possible pSB1A2+rbs+''dxs'' clone. (19.07.08)<br />
-
'''M49 to M54:''' pSB1A2-rbs+''crtI''<br />
+
'''M49 to M54:''' pSB1A2-rbs+''crtI''. (29.07.08: CF)<br />
-
'''M55 to M60:''' PZntA promoter<br />
+
'''M55 to M60:''' Putative pSB1A2+P''zntA'' (L23). (29.07.08: Yan, Nimisha) - M50 sequenced (31.07.08: CF)<br />
-
'''M61 to M66:''' BABEL2+rbs+''crtE''<br />
+
'''M61 to M66:''' Putative pSB1A2+rbs+''crtE'' (L22). (29.07.08: Yan, Nimisha) - M63 sequenced (31.07.08: CF)<br />
-
'''M67:''' rbs+''crtB''<br />
+
'''M67:''' rbs+''crtB'' (L21) (29.07.08: Yan, Nimisha) - Sequenced (31.07.08: CF)<br />
-
'''M68 to M71:''' pSB1A2+''glgC''-mut1,2 (31.07.08: AM, YAN)<br />
+
'''M68 to M71:''' pSB1A2+''glgC''-mut1,2 from plate 73 (31.07.08: AM, Yan)<br />
-
'''M72 to M75:''' pSB1A2+rbs+''dxs'' (31.07.08: AM, YAN)<br />
+
'''M72 to M75:''' pSB1A2+rbs+''dxs'' from plate 74 (31.07.08: AM, Yan)<br />
-
'''M76 to M79:''' BABEL2+''crtE'' (31.07.08: AM, YAN)<br />
+
'''M76 to M79:''' BABEL2+''crtE'' from plate 74 (31.07.08: AM, Yan)<br/>
-
'''M80 to M83:''' pSB1A2+rbs+''appY'' (04/08/08: YAN and OG)<br />
+
'''M80 to M83:''' pSB1A2+rbs+''appY'' from plate 83. (04.08.08: Yan, OG)<br />
-
'''M84 to M85:''' pSB1A2+rbs+''crtB'' (04/08/08: YAN and OG)<br />
+
'''M84 to M85:''' pSB1A2+rbs+''crtB'' from plate 84. (04.08.08: Yan, OG)<br />
-
'''M86 to M90:''' pSB1A2+''cenA'' (04/08/08: YAN and OG)<br />
+
'''M86 to M90:''' pSB1A2+''cenA'' from plate 85. (04.08.08: Yan, OG)<br />
-
'''M91 to M96:''' pSB1A2+''cex''(04/08/08: YAN and OG)<br />
+
'''M91 to M96:''' pSB1A2+''cex'' from plate 86. (04.08.08: Yan, OG)<br />
-
'''M97 to M104:''' pSB1A2+''cenA'' (05.08.08: YAN)<br />
+
'''M97 to M104:''' pSB1A2+''cenA'' from plate 85. (05.08.08: Yan)<br />
-
'''M105 to M108:''' pSB1A2+''cex'' (05.08.08: YAN)<br />
+
'''M105 to M108:''' pSB1A2+''cex'' from plate 85. (05.08.08: Yan)<br />
 +
'''M109 to M112:''' pSB1A2+''crtB''+''crtI'' and plate 102 (11.08.08: OG)<br />
 +
'''M113 to M116:''' pSB1A2+''glgC''-mut1,2 and plate 103 (11.08.08: OG)<br />
 +
'''M117 to M120:''' pSB1A2+''glgC''-mut1,2,3 and plate 94 (11.08.08: OG)<br />
 +
'''M121 to M123:''' pSB1A2-''crtY'' (12.08.08: OG)<br />
 +
'''M124 to M126:''' pSBA12-rbs+''crtY'' (12.08.08: OG)<br />
 +
'''M127 to M130:''' pSB1A2-P''cstA'' from plate 115 (13.08.08: OG)<br />
 +
'''M131 to M134:''' Minipreps of ''dxs+crtE'' (1,2,3,4) Note: M132 might be M136 (13.08.08: OG)<br />
 +
'''M135 to M138:''' Minipreps of ''dxs+LIMS1'' (1,2,3,4) Note: M136 might be M132 (13.08.08: OG)<br />
 +
'''M139 to M143:''' Minipreps of pSB1A2 +''crtE'' (1 to 4) Note: M142 and M143 both made from culture 4.(15.08.08: YAN/OG)<br />
 +
'''M144 to M147:''' pSB1A2+''cenA'' from plate 129 - Methanol used in preparation rather than ethanol! (18.08.08: OG)<br />
 +
'''M148 to M151:''' pSB1A2+''cex'' from plate 129. (18.08.08: OG)<br />
 +
'''M152 to M155:''' pSB1A2+''dxs+crtE'' from plate 130 (18.08.08: OG)<br />
 +
'''M156 to M159:''' pSB1A2+''cenA'' (20.08.08: OG)<br/>
 +
'''M160 to M163:''' Plate 155 streaks 1-4 (L48, pSB1A2+''dxs+lims+appY'') (25.08.08: AH)<br/>
 +
'''M164''': Plate 155 streak 5 (L49, pSB1A2+''crtB+crtI+appY'') (25.08.08: AH)<br/>
 +
'''M165 to M168:''' Plate 158 streaks 1-4 (L50, SOB+''glgC''-mut1,2) (25.08.08: AH)<br/>
 +
'''M169 to M172:''' Plate 159 streaks 1-4 (L51, SOB+''glgC''-mut1,2,3) (25.08.08: AH, Yan)<br/>
 +
'''M173:''' Plate 160 streak 1 (L55 (SOB+rbs+''glgC''-mut1,2,3) (25.08.08: Yan)<br/>
 +
'''M174 to M175:''' Plate 160 streaks 2 and 3 (L52, SOB+''glgC''-mut1,2,3) (25.08.08: Yan)<br/>
 +
'''M176 to M178:''' Plate 161 streaks 1-3 (L53, SOB+rbs+glgC-mut1,2) (25.08.08: Yan)<br/>
 +
'''M179 to M181:''' Plate 163 streaks 1-3 (L56, pSB1A2+p''cstA''+''lacZ' '') (25.08.08: Yan)<br/>
 +
'''M182 to M187:''' pSB1A2+''cenA'' (01.09.08: YAN)<br/>
 +
'''M188 to M193:''' ''crtBI+appY'' (01.09.08: YAN)<br/>
 +
'''M194 to M197:''' ''appY+dxs+LIMS1'' (02.09.08:YAN)<br/>
 +
'''M198 to M200:''' L58 = self-ligation of P91 (SOB2-''SU1''). M199 and M200 were missed out by accident. (09.09.08: Yan)<br/>
 +
'''M201 to M203:''' L59 = self-ligation of P92 (SOB2-rbs-''SU1''). (09.09.08: Yan)<br/>
 +
'''M204 to M206:''' L60 = self-ligation of P93 (SOB2-''ISO2''). (09.09.08: Yan)<br/>
 +
'''M207 to M209:''' L61 = self-ligation of P94 (SOB2-rbs-''ISO2''). (09.09.08: Yan)<br/>
 +
'''M210 to M212:''' pSB1A2-''glgc'' triple mutants. (15.09.08: AM)<br/>
 +
'''M213:''' ''dxs+crtEBI''. (15.09.08: AM)<br/>
 +
'''M214:''' pSB1A2-''cenA''. (15.09.08: AM)<br/>
 +
'''M215 to M216:''' pSB1A2-''bglX''. (15.09.08: AM)<br/>
 +
'''M217 to M221:''' ''dxs+crtEBI''. (15.09.08: AM)<br/>
 +
'''M222 to M225:''' P''lac+dxs+LIMS1+appY'' patches 1, 2, 6 and 7 (CF, 23-9-8). All look OK on gel. M222 confirmed by sequencing.<br/>
 +
'''M226 to M229:''' SOB-''SU1''. (CF, 27-9-8) Only M229 looks plausible on gel, but sequencing shows problem at end. <br/>
 +
'''M230 to M232:''' SOB-''ISO2''. (CF, 27-9-8) All looked OK; sequencing showed M230 and 232 OK, 231 problem with suffix.<br/>
 +
'''M233 to M235:''' pSB1A2-''cex'' mutant (CF) Sequence showed M233 OK.<br/>
 +
'''M236 to M238:''' pSB1A2-''cenA'' (CF, 27-9-8) . M236 clearly wrong on gel, others plausible. Sequence showed M237 OK.<br/>
 +
'''M239 to M241:''' pSB1A2-''bglX'' (CF, 27-9-8) Only 240 looked plausible on gel, sequence showed wrong.<br/>
 +
'''M242 to M243:''' pSB1A2-rbs-''glgC'' (CF, 28-9-8)<br/>
 +
'''M244 to M245:''' pSB1A2-rbs-''glgC16'' (CF, 28-9-8)<br/>
 +
'''M246 to M247:''' pSB1A2-''bglX'' (CF, 28-9-8). M247 looks plausible but sequencing shows wrong. <br/>
 +
'''M248 to M253:''' pSB1A2-''crtBI-dxs-crtE'' (CF, 30-9-8). All look wrong.<br/>
 +
'''M254 to M255:''' pSB1A2-P''lac-glgC'' (CF, 2-10-8). Look OK. M254 confirmed by sequencing.<br/>
 +
'''M256 to M257:''' pSB1A2-P''lac-glgC16'' (CF, 2-10-8). Look OK. M256 confirmed by sequencing.<br/>
 +
'''M258:''' pSB1A2-P''cstA''-''lacZ' '' (CF, 2-10-8). Looks OK, confirmed by sequencing.<br/>
 +
'''M259:''' pSB1A2-P''cstA''-''xylE'' (CF, 2-10-8). Looks OK, confirmed by sequencing.<br/>
 +
'''M260 to M265:''' SOB-''SU1'' (CF, 3-10-8). M265 looks OK on gel, confirmed by sequencing.<br/>
 +
'''M266 to M269:''' SOB-''ISO2'' (CF, 4-10-8). M269 looks plausible on gel.<br/>
 +
'''M270 to M272:''' pSB1A2+rbs+''cex'' (CF, 8-10-8). All look wrong.<br/>
 +
'''M273 to M275:''' pSB1A2+rbs+''cenA'' (CF, 8-10-8). M273, 274 looked OK but turned out to be wrong gene.<br/>
 +
'''M276 to M278:''' pSB1A2+rbs+''cex'' (CF, 11-10-8). M276 looked OK but turned out to be wrong gene.<br/>
 +
'''M279 to M282:''' pSB1A2+''bglX'' (CF, 14-10-8). All looked Ok on gel, but sequencing showed that all but M281 had frameshift mutations in a run of A near the start resulting in premature termination. M281 seems OK though.<br/>
 +
'''M283 to M286:''' from mutagenesis of ''SU1'' (CF, 14-10-8 and 15-10-8). M286 looks wrong, others still seem to have the EcoRI site.<br/>
 +
'''M287 to M290:''' from mutagenesis of ''ISO2'' (CF, 15-10-8). All seem to still have EcoRI site.<br/>
 +
'''M291 to M293:''' pSB1A2+P''lac+crtBI'' (CF, 16-10-8). All look OK.<br/>
 +
'''M294 to M296:''' pSB1A2+P''lac+dxs.crtE'' (CF, 16-10-8). All look OK.<br/>
 +
'''M297 to M300:''' from mutagenesis of ''SU1'' (CF, 19-10-8). All seem to still have EcoRI site.<br/>
 +
'''M301:''' from mutagenesis of ''ISO2'' (CF, 19-10-8). Seems to still have EcoRI site.<br/>
 +
'''M302:''' supposed to be P''lac+dxs.crtEBI'' but seems to be just P''lac+dxs.crtE'' (CF, 19-10-8).<br/>
 +
'''M303 to M304:''' pSB1A2+rbs+''cex'' (CF, 26-10-8). Both look OK.<br/>
 +
'''M305 to M306:''' pSB1A2+rbs+''cenA'' (CF, 26-10-8). Both look OK.<br/>
 +
'''M307 to M308:''' pSB1A2+rbs+''bglX'' (CF, 26-10-8). Both look OK.<br/>

Latest revision as of 00:19, 30 October 2008

< Back to Notebook

Minipreps of Confirmed Clones

  • M2: pSB1A2+dxs
  • M15: pSB1A2+appY
  • (?) M36: pSB1A2+crtB (sequencing results not recorded, but follow-up did occur)
  • (?) M42: pSB1A2+crtI (sequencing results not recorded, but follow-up did occur)
  • M43: BABEL2(?)+glgC double mutant.
  • M50: pSB1A2+rbs+crtI
  • M63: BABEL2+rbs+crtE
  • M67: pSB1A2+rbs+crtB
  • M72: pSB1A2+rbs+dxs
  • M79: BABEL2+crtE
  • M82: pSB1A2+rbs+appY
  • M109: pSB1A2+crtBI
  • (?) M121: pSB1A2+crtY (sequencing results not posted; no follow-up recorded)
  • (?) M124 pSB1A2+rbs+crtY (sequencing results not posted; no follow-up recorded)
  • M116: pSB1A2+glgC double mutant
  • M130: pSB1A2+PcstA
  • M137: pSB1A2+dxs+LIMS1
  • M150: pSB1A2+cex
  • M152: pSB1A2+dxs+crtE
    • M154: not sequenced, but similar to M152.
    • M155: not sequenced, but also similar to M152; ligated to M110 (crtBI) in L62.
  • M162: pSB1A2+dxs+LIMS1+appY
  • M212: pSB1A2+glgC triple mutant (ie. glgC16)
  • M222: pSB1A2+Plac+dxs+LIMS1+appY
  • M233: pSB1A2+cex mutant (PstI site removed)
    • M234, 235: not sequenced, but similar to M233.
  • M237, 238: pSB1A2+cenA
  • M243: pSB1A2+rbs+glgC
  • M245: pSB1A2+rbs+glgC16
  • M254: pSB1A2+Plac+rbs+glgC
  • M256: pSB1A2+Plac+rbs+glgC16
  • M258: pSB1A2+PcstA+lacZ' (BBa_J15202)
  • M259: pSB1A2+PcstA+xylE (BBa_J33204)
  • M281: pSB1A2+bglX coding sequence
  • M303: pSB1A2+rbs+cex
  • M305: pSB1A2+rbs+cenA
  • M307: pSB1A2+rbs+bglX

Complete List of Minipreps

M1 to M6: pSB1A2+dxs transformants (plate 11). M2, M3, M4 and M6 all looked correct on gel 2. (30.06.08) - M2 was sequenced to confirm identity and was maxiprepped as X2.
M7: BABEL1+glgC transformant (plate 12). Did not look right on gel. (30.06.08)
M8 to M12: BABEL2+glgC transformants (plate 13). M10 and M11 both looked good on a gel, but sequencing showed that both had the insert in the reverse orientation. (30.06.08)
M13 to M18: pSB1A2+appY transformants. Gel 10 showed that M15 to M18, and possibly M14, all seemed to have inserts the right size. (09.07.08)
M19 to M24: BABEL2+glgC clones after mutation of EcoRI site 2. EcoRI digests on *Gel 11* suggest that *M19, M21 and M22* may have lost the EcoRI site. (10.07.08)
M25 to M30: possible rbs+dxs transformants from plate 34. (13.07.08)
M31 to M36: pSB1A2+crtB clones from plate 35. (14.07.08)
M37 to M42: pSB1A2+crtI clones from plate 36. (14.07.08)
M43 to M47: possible BABEL2+glgC double mutants (both EcoRI sites gone). (19.07.08)
M48: a possible pSB1A2+rbs+dxs clone. (19.07.08)
M49 to M54: pSB1A2-rbs+crtI. (29.07.08: CF)
M55 to M60: Putative pSB1A2+PzntA (L23). (29.07.08: Yan, Nimisha) - M50 sequenced (31.07.08: CF)
M61 to M66: Putative pSB1A2+rbs+crtE (L22). (29.07.08: Yan, Nimisha) - M63 sequenced (31.07.08: CF)
M67: rbs+crtB (L21) (29.07.08: Yan, Nimisha) - Sequenced (31.07.08: CF)
M68 to M71: pSB1A2+glgC-mut1,2 from plate 73 (31.07.08: AM, Yan)
M72 to M75: pSB1A2+rbs+dxs from plate 74 (31.07.08: AM, Yan)
M76 to M79: BABEL2+crtE from plate 74 (31.07.08: AM, Yan)
M80 to M83: pSB1A2+rbs+appY from plate 83. (04.08.08: Yan, OG)
M84 to M85: pSB1A2+rbs+crtB from plate 84. (04.08.08: Yan, OG)
M86 to M90: pSB1A2+cenA from plate 85. (04.08.08: Yan, OG)
M91 to M96: pSB1A2+cex from plate 86. (04.08.08: Yan, OG)
M97 to M104: pSB1A2+cenA from plate 85. (05.08.08: Yan)
M105 to M108: pSB1A2+cex from plate 85. (05.08.08: Yan)
M109 to M112: pSB1A2+crtB+crtI and plate 102 (11.08.08: OG)
M113 to M116: pSB1A2+glgC-mut1,2 and plate 103 (11.08.08: OG)
M117 to M120: pSB1A2+glgC-mut1,2,3 and plate 94 (11.08.08: OG)
M121 to M123: pSB1A2-crtY (12.08.08: OG)
M124 to M126: pSBA12-rbs+crtY (12.08.08: OG)
M127 to M130: pSB1A2-PcstA from plate 115 (13.08.08: OG)
M131 to M134: Minipreps of dxs+crtE (1,2,3,4) Note: M132 might be M136 (13.08.08: OG)
M135 to M138: Minipreps of dxs+LIMS1 (1,2,3,4) Note: M136 might be M132 (13.08.08: OG)
M139 to M143: Minipreps of pSB1A2 +crtE (1 to 4) Note: M142 and M143 both made from culture 4.(15.08.08: YAN/OG)
M144 to M147: pSB1A2+cenA from plate 129 - Methanol used in preparation rather than ethanol! (18.08.08: OG)
M148 to M151: pSB1A2+cex from plate 129. (18.08.08: OG)
M152 to M155: pSB1A2+dxs+crtE from plate 130 (18.08.08: OG)
M156 to M159: pSB1A2+cenA (20.08.08: OG)
M160 to M163: Plate 155 streaks 1-4 (L48, pSB1A2+dxs+lims+appY) (25.08.08: AH)
M164: Plate 155 streak 5 (L49, pSB1A2+crtB+crtI+appY) (25.08.08: AH)
M165 to M168: Plate 158 streaks 1-4 (L50, SOB+glgC-mut1,2) (25.08.08: AH)
M169 to M172: Plate 159 streaks 1-4 (L51, SOB+glgC-mut1,2,3) (25.08.08: AH, Yan)
M173: Plate 160 streak 1 (L55 (SOB+rbs+glgC-mut1,2,3) (25.08.08: Yan)
M174 to M175: Plate 160 streaks 2 and 3 (L52, SOB+glgC-mut1,2,3) (25.08.08: Yan)
M176 to M178: Plate 161 streaks 1-3 (L53, SOB+rbs+glgC-mut1,2) (25.08.08: Yan)
M179 to M181: Plate 163 streaks 1-3 (L56, pSB1A2+pcstA+lacZ' ) (25.08.08: Yan)
M182 to M187: pSB1A2+cenA (01.09.08: YAN)
M188 to M193: crtBI+appY (01.09.08: YAN)
M194 to M197: appY+dxs+LIMS1 (02.09.08:YAN)
M198 to M200: L58 = self-ligation of P91 (SOB2-SU1). M199 and M200 were missed out by accident. (09.09.08: Yan)
M201 to M203: L59 = self-ligation of P92 (SOB2-rbs-SU1). (09.09.08: Yan)
M204 to M206: L60 = self-ligation of P93 (SOB2-ISO2). (09.09.08: Yan)
M207 to M209: L61 = self-ligation of P94 (SOB2-rbs-ISO2). (09.09.08: Yan)
M210 to M212: pSB1A2-glgc triple mutants. (15.09.08: AM)
M213: dxs+crtEBI. (15.09.08: AM)
M214: pSB1A2-cenA. (15.09.08: AM)
M215 to M216: pSB1A2-bglX. (15.09.08: AM)
M217 to M221: dxs+crtEBI. (15.09.08: AM)
M222 to M225: Plac+dxs+LIMS1+appY patches 1, 2, 6 and 7 (CF, 23-9-8). All look OK on gel. M222 confirmed by sequencing.
M226 to M229: SOB-SU1. (CF, 27-9-8) Only M229 looks plausible on gel, but sequencing shows problem at end.
M230 to M232: SOB-ISO2. (CF, 27-9-8) All looked OK; sequencing showed M230 and 232 OK, 231 problem with suffix.
M233 to M235: pSB1A2-cex mutant (CF) Sequence showed M233 OK.
M236 to M238: pSB1A2-cenA (CF, 27-9-8) . M236 clearly wrong on gel, others plausible. Sequence showed M237 OK.
M239 to M241: pSB1A2-bglX (CF, 27-9-8) Only 240 looked plausible on gel, sequence showed wrong.
M242 to M243: pSB1A2-rbs-glgC (CF, 28-9-8)
M244 to M245: pSB1A2-rbs-glgC16 (CF, 28-9-8)
M246 to M247: pSB1A2-bglX (CF, 28-9-8). M247 looks plausible but sequencing shows wrong.
M248 to M253: pSB1A2-crtBI-dxs-crtE (CF, 30-9-8). All look wrong.
M254 to M255: pSB1A2-Plac-glgC (CF, 2-10-8). Look OK. M254 confirmed by sequencing.
M256 to M257: pSB1A2-Plac-glgC16 (CF, 2-10-8). Look OK. M256 confirmed by sequencing.
M258: pSB1A2-PcstA-lacZ' (CF, 2-10-8). Looks OK, confirmed by sequencing.
M259: pSB1A2-PcstA-xylE (CF, 2-10-8). Looks OK, confirmed by sequencing.
M260 to M265: SOB-SU1 (CF, 3-10-8). M265 looks OK on gel, confirmed by sequencing.
M266 to M269: SOB-ISO2 (CF, 4-10-8). M269 looks plausible on gel.
M270 to M272: pSB1A2+rbs+cex (CF, 8-10-8). All look wrong.
M273 to M275: pSB1A2+rbs+cenA (CF, 8-10-8). M273, 274 looked OK but turned out to be wrong gene.
M276 to M278: pSB1A2+rbs+cex (CF, 11-10-8). M276 looked OK but turned out to be wrong gene.
M279 to M282: pSB1A2+bglX (CF, 14-10-8). All looked Ok on gel, but sequencing showed that all but M281 had frameshift mutations in a run of A near the start resulting in premature termination. M281 seems OK though.
M283 to M286: from mutagenesis of SU1 (CF, 14-10-8 and 15-10-8). M286 looks wrong, others still seem to have the EcoRI site.
M287 to M290: from mutagenesis of ISO2 (CF, 15-10-8). All seem to still have EcoRI site.
M291 to M293: pSB1A2+Plac+crtBI (CF, 16-10-8). All look OK.
M294 to M296: pSB1A2+Plac+dxs.crtE (CF, 16-10-8). All look OK.
M297 to M300: from mutagenesis of SU1 (CF, 19-10-8). All seem to still have EcoRI site.
M301: from mutagenesis of ISO2 (CF, 19-10-8). Seems to still have EcoRI site.
M302: supposed to be Plac+dxs.crtEBI but seems to be just Plac+dxs.crtE (CF, 19-10-8).
M303 to M304: pSB1A2+rbs+cex (CF, 26-10-8). Both look OK.
M305 to M306: pSB1A2+rbs+cenA (CF, 26-10-8). Both look OK.
M307 to M308: pSB1A2+rbs+bglX (CF, 26-10-8). Both look OK.

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