Glasgow/Wetlab/Week8

From 2007.igem.org

(Difference between revisions)
(Tuesday 21st August 2007)
(Tuesday 21st August 2007)
Line 25: Line 25:
<ol>
<ol>
<li>
<li>
-
 
+
Christine cloned (*a→g*) into TOPO vectors using different ratios of PCR product:TOPO vector (see [[Glasgow/Wetlab/Protocols#Protocol 12: TOPO Cloning Reaction|Protocol 12]] ).  Ratios were 0.5ul:1ul, 1ul:1ul, and 2ul:1ul.
 +
<li>
 +
TOPO vectors with different ratios of PCR product:TOPO vector were tranformed (see [[Glasgow/Wetlab/Protocols#Protocol 13: One Shot Chemical Transformation Protocol for Competent E. coli Reaction|Protocol 13]] (with 30 sec heat shock at 42°C and 250ul of 37°C LB instead of SOC medium))and grown overnight on kanamycin plates at 37°C, each reaction divided into 100ul and ~200ul per plate.
<li>
<li>
Maija digested the construction vectors 3/20G and 4/6B, and the 7 gene operon with SpeI and XbaI ([[Glasgow/Wetlab/Protocols#Protocol 7: Restriction Digests|Protocol 7]]) with the intention of then ligating the 7 gene operon into the construction vectors.  The 7 gene operon DNA (taken from sample 2 of the PCR which was gel extracted 13/08/07) did not appear on the gel.
Maija digested the construction vectors 3/20G and 4/6B, and the 7 gene operon with SpeI and XbaI ([[Glasgow/Wetlab/Protocols#Protocol 7: Restriction Digests|Protocol 7]]) with the intention of then ligating the 7 gene operon into the construction vectors.  The 7 gene operon DNA (taken from sample 2 of the PCR which was gel extracted 13/08/07) did not appear on the gel.

Revision as of 18:08, 21 August 2007

Back To Main Page | Back To Wetlab Log


Contents

Week 8

Monday 20th August 2007

  1. Chrsitine did more minipreps for colonies suspected of containing the 7 gene operon. When these minipreps were run on a gel there was no evidence of an insert.


  1. These
  2. are
    1. numbered
  3. sections



This is a table
Look at the code to see the template.

Tuesday 21st August 2007

  1. Christine cloned (*a→g*) into TOPO vectors using different ratios of PCR product:TOPO vector (see Protocol 12 ). Ratios were 0.5ul:1ul, 1ul:1ul, and 2ul:1ul.
  2. TOPO vectors with different ratios of PCR product:TOPO vector were tranformed (see Protocol 13 (with 30 sec heat shock at 42°C and 250ul of 37°C LB instead of SOC medium))and grown overnight on kanamycin plates at 37°C, each reaction divided into 100ul and ~200ul per plate.
  3. Maija digested the construction vectors 3/20G and 4/6B, and the 7 gene operon with SpeI and XbaI (Protocol 7) with the intention of then ligating the 7 gene operon into the construction vectors. The 7 gene operon DNA (taken from sample 2 of the PCR which was gel extracted 13/08/07) did not appear on the gel.
    Label Description Enzymes Expected size Result
    3/20G Construction vector SpeI, XbaI 2071bp 2071bp
    4/6B Construction vector SpeI, XbaI 2928bp 2928bp
    (*a→g*) 7 gene operon SpeI, XbaI 6263bp No result
  4. Christine gel extracted 4/6B and 3/20G (see Protocol 11) using 30ul ddH2O at 60°C to elute the DNA.

    Wednesday 22nd August 2007

    Thursday 23rd August 2007

    === Friday 24th August 2007 ===