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Team:SDU-Denmark/labnotes9
From 2010.igem.org
Lab notes (9/6 - 9/12)
Contents |
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
| 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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