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| - | Following digestion, the samples were loaded and run on a 1.5% agarose gel, before being extracted prior to ligation. | + | Following digestion, the samples were loaded and run on a 1.5% agarose gel, before being extracted prior to ligation.<br> |
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Revision as of 18:09, 12 September 2010
Lab notes (9/6 - 9/12)
Flagella Group
Restriction Digest of Gel Extracted FlhDCmut, pSB1C3 and pSB1AK3
Date: 9/12 2010
Done By: Sheila
Protocol: RD1.1
Notes: The restriction digests are made in preperation for two different ligations, the first is the ligation of FlhDCmut into the submission backbone; pSB1C3. The second is the the ligation of FlhDCmut into pSB1AK3. Different restriction enzymes were used in, as shown in the following tables
Restriction Digest mixtures
FlhDCmut and pSB1C3
Restriction digest
FlhDCmut
| H2O |
48μL |
| EcoR1 |
4μL |
| Pst1 |
4μL |
| FD Green buffer |
8μL |
| Sample |
20μL |
pSB1C3
| H2O |
24μL |
| EcoR1 |
2μL |
| Pst1 |
2μL |
| FD Green buffer |
4μL |
| Sample |
10μL |
FlhDCmut and pSB1AK3
Restriction digest
FlhDCmut
| H2O |
48μL |
| EcoR1 |
4μL |
| Spe1 |
4μL |
| FD Green buffer |
8μL |
| Sample |
20μL |
pSB1AK3
| H2O |
24μL |
| EcoR1 |
2μL |
| Xba1 |
2μL |
| FD Green buffer |
4μL |
| Sample |
10μL |
Following digestion, the samples were loaded and run on a 1.5% agarose gel, before being extracted prior to ligation.
Results:
Ligation of FlhDCmut and pSB1C3
Date: 9/12 2010
Done By: Sheila
Protocol: LG1.2
Notes:
Results:
Ligation of FlhDCmut and pSB1AK3
Date: 9/12 2010
Done By: Sheila
Protocol: LG1.2
Notes:
Results:
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