Glasgow/Wetlab/Week8
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| - | + | 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. | |
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| + | 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. | ||
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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
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Contents |
Week 8
Monday 20th August 2007
- 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.
- These
- are
- numbered
- sections
| This is | a table |
| Look at the code | to see the template. |
Tuesday 21st August 2007
- 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.
- 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.
-
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 -
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 ===
