Team:Johns Hopkins/Notebook

From 2008.igem.org

(Difference between revisions)
(GROUP 1: Fluorescent Proteins)
(GROUP 1: Fluorescent Proteins)
Line 45: Line 45:
   Part no.: BBa_K110017 -> BBa_K110023
   Part no.: BBa_K110017 -> BBa_K110023
   Part Description: yESapphire , mCherry, venusYFP, and Citrine <br>
   Part Description: yESapphire , mCherry, venusYFP, and Citrine <br>
-
   <p>Work on yESapphire was gratiously done by James. Primers were designed. Restriction site ends have been added through PCR. The
+
   Work on yESapphire was gratiously done by James. Primers were designed. Restriction site ends have been added through PCR. The
   product was cloned into JM109. Colonies were picked and an inoculation was grown. Miniprep was performed and subsequent DNA
   product was cloned into JM109. Colonies were picked and an inoculation was grown. Miniprep was performed and subsequent DNA
   was CS PCR'd and run on a gel to verify contents <b>([http://www.jhu.edu/iGEM/Group3:ShortTwo-wayStops/2008-7-22.Short%202Way%20Stop,%20Alpha%20Promoters,%20&%20Sapphire%20FP.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html  Short 2Way Stop, Alpha Promoters, & Sapphire FP])</b>. BioBrick is currently being
   was CS PCR'd and run on a gel to verify contents <b>([http://www.jhu.edu/iGEM/Group3:ShortTwo-wayStops/2008-7-22.Short%202Way%20Stop,%20Alpha%20Promoters,%20&%20Sapphire%20FP.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html  Short 2Way Stop, Alpha Promoters, & Sapphire FP])</b>. BioBrick is currently being

Revision as of 03:47, 1 August 2008

Contents

Groups

iGEM Groups 1.0
iGEM Groups 2.01
iGEM Groups 2.02

Important reminders and notes

 [Can make general comments here, so they don't get lost in peoples e-mail boxes]
 
 July 11: Primers for group 1 were delivered yesterday.
 July 11: Lab meeting at 7:30PM in the lab to go over miniprep protocol.
 July 15: Lab meeting at 6:30PM with Jessica. Have status reports ready. 
          Bring labtop if you can.
 July 17: Restriction Digest/Sequencing Preparation (with James) 6:00PM.
 July 21: 6:00PM or 6:30PM Lab meeting with Jessica. Have status reports ready.
 July 29: 7:00PM Lab Meeting- Meet in Conference Room across from lab. Have Status Reports ready

Data

To upload data, go [http://www.jhu.edu/iGEM/X_files/Read2.html here], click on [http://www.jhu.edu/iGEM/X_files/Read2.html upload data], and provide the necessary information and results.

How to submit data:

1. log-in as you once had to from the www.jhu.edu/iGEM website "login"

  • User: ****** etc...
  • Pass: ***** etc...

2. click on UPLOAD DATA from the 'x-files page'
3. add data etc.... and click submit: This generates a webpage and the URL to it is linked in the page you are directed to after you press submit. Copy that URL and past it into the wiki or into the web-browser url box to see what it looks like.
\* If you find that the picture you are uploading is not showing up e-mail Tejas.

Status Reports

The status reports of each group below will continuously be updated as we work on the biobricks. The following PDFs contain progressive versions of our status reports as we continue through the sex detector project; they are added weekly. To learn more about each biobrick, please refer to the Biobrick page.

Status Report 2.1 - 07/12/08
Status Report 2.2 - 07/17/08

Please BOLD the most recent step that you have completed. Do this by placing the tag < b > in front of and </ b > at the end of what you would like to be placed in bold (with no space between letter and carrot symbol (<,>). Click on your group name for the detailed changes occurring to each biobrick.

GROUP 1: Fluorescent Proteins

Summary for Fluorescent Proteins Group

 Date: July 22, 2008
 Status report by: Tejas, Ingrid
 Part no.: BBa_K110017 -> BBa_K110023
 Part Description: yESapphire , mCherry, venusYFP, and Citrine 
Work on yESapphire was gratiously done by James. Primers were designed. Restriction site ends have been added through PCR. The product was cloned into JM109. Colonies were picked and an inoculation was grown. Miniprep was performed and subsequent DNA was CS PCR'd and run on a gel to verify contents ([http://www.jhu.edu/iGEM/Group3:ShortTwo-wayStops/2008-7-22.Short%202Way%20Stop,%20Alpha%20Promoters,%20&%20Sapphire%20FP.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html Short 2Way Stop, Alpha Promoters, & Sapphire FP]). BioBrick is currently being sequenced.
</p> Work on mCherry and venusYFP (BBa_K110018 -> BBa_K110021) is currently being done. Primers were designed. Restriction sites have been added through PCR. ([http://www.jhu.edu/iGEM/Group1:FluorescentProteins/2008-7-23.PCR%20Products:%20mCherry%20and%20Venus%20YFP,%20both%20RtL%20and%20LtR.Ingrid,%20Tejas.html PCR Products: mCherry and Venus YFP, both RtL and LtR]) The products were cloned into JM109 and plated. 3 colonies per 4 BioBricks (total 12) were picked, grown out, miniprepped, and digested for verification on a gel.
According to the results, ([http://www.jhu.edu/iGEM/Group1:FluorescentProteins/2008-7-22.Venus%20YFP%20and%20mCherry%20Miniprep%20check%20via%20digest.Ingrid.html Venus YFP and mCherry Miniprep check via digest]) , only one of the mCherry's (BBa_K110019) is the correct product. A second Digest was preformed to check this, meanwhile new colonies have been picked on 7.22.2008 and are being grown out for a higher yield of DNA so that it can be sent off for sequencing. [http://www.jhu.edu/iGEM/Group1:FluorescentProteins/2008-7-23.Re-Digest%20of%20three%20samples%20of%20mCherry%20BBa_K110018.Ingrid%20.html Re-Digest of three samples of mCherry BBa_K110018]
Work on Citrine has not yet begun. Primers have been designed and ordered, but template DNA must be grown out. Template DNA will be grown and extracted by James should we later decide that Citrine will be needed.
*Note that mCherry and Citrine both have restriction sites within the coding region, and are therefore not optimal. Advice/help on this issue would be appreciated.

GROUP 2: MATa Specific-promoters

Summary for MATa Specific Promoters Group

 Date: July 29, 2008
 Status report by: Allison and Nate
 Part no.: BBa_K110008 -> BBa_K110016
 Part Description: A promoters: MFA1 (L+R) and Ste2 (R+L), respectively.
 Two samples of mini preps from MFA1 and Ste2 were sent off for sequencing at the end of last week.
 Results will follow soon.

GROUP 3: Short Two-Way Stops

Summary for Short Two-Way Stops Group

 Date 7/29/08
 Status report by: James
 Part no.: BBa_K110011 -> BBa_K110013; BBa_K110010;17
 Part Description: Short two way stops
 Summary: Sequencing of BBa_K110010;17 (Sapphire FP) revealed confusing, if not bad
 sequences.Sequence Data  
 PCR cloning and transformation was completed sucessfully last week on the Short two way stops
 A Miniprep was done and a  RE digest will be completed soon (possibly tonight)


 Date: 7/22/2008
 Status report by: James
 Part no.: BBa_K110011 -> BBa_K110013
 Part Description: Short two way stops
 Summary: RE digestion showed no insert the plasmid of our clones, thus the PCR was done again.
 [http://www.jhu.edu/iGEM/Group3:ShortTwo-wayStops/2008-7-23.Short%20way%20stops.James.html Short way stops]

GROUP 4: Long Two-Way Stops & MAT(alpha) Specific Promotors

Summary for Long Two-Way Stops & MAT(alpha) Specific Promoters Group

 Date: July 29, 2008
 Status report by: Jaime
 Part no.: BBa_K110001, BBa_K110003, BBa_K110005, BBa_K110006
 Part Description: Long Two-way Stops & Mat(alpha) specific promotors
 [http://www.jhu.edu/iGEM/Group4:LongTwo-wayStopsANDMATalphaSpecificPromoters/2008-7-25.Restriction%20Enzyme%20Digest%20of%20Mini-Preps.Jaime.html Restriction Enzyme Digest of Mini-Preps]
 
 Summary here:
 Need to be sequenced.

GROUP 5: MATa Specific Promoters II

Summary for MATa Specific Promoters II Group

 Date: _________ __, 2008
 Status report by: _____
 Part no.: BBa_K1100XX -> BBa_K1100YY
 Part Description:
 
 Summary here.

GROUP 6: Vectors

Summary for Vectors Group

 Date: July 31, 2008
 Status report by: Tejas
 Part no.: BBa_K1100XX -> BBa_K1100YY
 Part Description: Vectors for concatenating and executing BioBricks
 
 The vectors to be used were delivered to us in STABS from MIT. They are pSB4A5 (amp), pSB4C5 (cam), pSB3K5 (kan), and pSB4K5 (kan).
 The vectors come pre-inserted with the ccdB gene, preventing growth in all E. coli strains except DB3.1. To produce sufficient
 quantities of thee vectors, I grew out the strains and performed several minipreps. Total amount of DNA is estimated between
 5 to 20 ug per vector. Restriction digests confirmed the extracted DNA was vectors.
 Transformation into DB3.1 competent cells was unsuccessful, likely due to the possibility that the competent cells may not
 actually be DB3.1. Currently making new competent cells from old DB3.1 strain (BB #5777) and from new DB3.1 strain (BB #2198).

GROUP 7: Microscopy/Yeast

Summary for Microscopy/Yeast Group

 Date: _________ __, 2008
 Status report by: _____
 Part no.: BBa_K1100XX -> BBa_K1100YY
 Part Description:
 
 Summary here.