Team:Warsaw/Calendar-Main/6 October 2008

From 2008.igem.org

(Difference between revisions)

Revision as of 20:38, 28 October 2008


	Attribution analyzer / Interactive list of topics

Preparation of linker_alpha (BBa_K103009)

Michał K.

Colony PCR with LinLSXNE and AlphaPSpe primers on colonies from plates with transformations pSB1A3+ linker_alpha (BBa_K103009) (annealing temperature - 55°C,60 s of elongation step). No visible bands for proper product (Fig. 1).
Inoculation of some colonies which grown on plate - pSB2K3 + linker_alpha (BBa_K103009) to liquid LB + kanamycin.

Fig. 1.Colony PCR with LinLSXNE and AlphaPSpe. Lack of expected 400 bp product.(Kolonijny_alfa.jpg

Preparation of linker_omega (BBa_K103013)

Michał K.

Inoculation of some colonies which grown on plate - pSB2K3 + linker_omega (BBa_K103013) to liquid LB + kanamycin.

Preparation of OmpA-linker-omega-linker (BBa_K103016)

Michał K.

Colony PCR with OmpaL_N and OmpaP_link primers on colonies from plates with transformations pACYC177 + OmpA-linker-omega-linker (BBa_K103016) (annealing temperature - 55°C,90 s of elongation step). No visible bands for proper product (Fig. 2).
Inoculation of some colonies which grown on plate transformation: pACYC177 + OmpA-linker-omega-linker (BBa_K103016) to liquid LB + kanamycin.

Fig. 2.Colony PCR with OmpaL_N and OmpaP_link. Lack of expected 350 bp product.

Preparation of vector for pT7 constructs

Michał K.

Overnight selfligation of pET15b+OmpA_omega (with removed XbaI).

Preparation of OmpA_linker_omega_linker under Plac (BBa_K103018)

Michał K.

Overnight ligation of isolated DNA fragments pSB2K3 (from 1 October) + OmpA_linker_omega_linker under Plac (BBa_K103018) (without EcoRI site).

Preparation of AID(BBa_K103001)

Michał K.

Digest of pSB1A3 plasmid with XbaI and PstI (Tango buffer). Dephosphorylation (CIAP) of plasmid.
Gel electrophoresis and gel-out of proper band - 2200 bp. Fig. 3.

Fig. 3.Digest of pSB1A3. 1. Marker 2. pSB1A3

@@ Line 12: / Line 12: @@
 <ol><li> Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> with <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinLSXNE">LinLSXNE</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AlphaPSpe">AlphaPSpe</a>
-  primers on colonies from plates with transformations <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+ <a href=http://partsregistry.org/Part:BBa_K103009>linker_alpha (BBa_K103009)</a> (annealing temperature - 55&deg;C,60 s of elongation step). No visible bands for proper product (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/6_October_2008#fig1">Fig. 1.</a>).</li>
+  primers on colonies from plates with transformations <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a>+ <a href=http://partsregistry.org/Part:BBa_K103009>linker_alpha (BBa_K103009)</a> (annealing temperature - 55&deg;C,60 s of elongation step). No visible bands for proper product (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/6_October_2008#fig1">Fig. 1</a>).</li>
 <li>Inoculation of some colonies which grown on plate - <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/Part:BBa_K103009>linker_alpha (BBa_K103009)</a> to liquid LB + kanamycin. </li></ol>
@@ Line 34: / Line 34: @@
 <li> Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> with <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaL_N">OmpaL_N</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#OmpaP_link">OmpaP_link</a>
-  primers on colonies from plates with transformations <a href=http://partsregistry.org/Part:BBa_I739204>pACYC177</a> + <a href=http://partsregistry.org/Part:BBa_K103016>OmpA-linker-omega-linker (BBa_K103016)</a> (annealing temperature - 55&deg;C,90 s of elongation step). No visible bands for proper product (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/6_October_2008#fig2">Fig. 2.</a>).</li>
+  primers on colonies from plates with transformations <a href=http://partsregistry.org/Part:BBa_I739204>pACYC177</a> + <a href=http://partsregistry.org/Part:BBa_K103016>OmpA-linker-omega-linker (BBa_K103016)</a> (annealing temperature - 55&deg;C,90 s of elongation step). No visible bands for proper product (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/6_October_2008#fig2">Fig. 2</a>).</li>
 <li>Inoculation of some colonies which grown on plate transformation: <a href=http://partsregistry.org/Part:BBa_I739204>pACYC177</a> + <a href=http://partsregistry.org/Part:BBa_K103016>OmpA-linker-omega-linker (BBa_K103016)</a> to liquid LB + kanamycin. </li></ol>
@@ Line 59: / Line 59: @@
 <ol>
 <li> <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> plasmid with XbaI and PstI (Tango buffer). <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">Dephosphorylation</a> (CIAP) of plasmid.</li>
-<li> Gel electrophoresis (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/6_October_2008#fig3">Fig. 3.</a>) and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 2200 bp.</li>
+<li> Gel electrophoresis and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 2200 bp. <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/6_October_2008#fig3">Fig. 3</a>.</li>
 </ol>