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Team:Stockholm/16 September 2010
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====Colony PCR==== | ====Colony PCR==== | ||
Picked new colonies for colony PCR from 14/9 plates: | Picked new colonies for colony PCR from 14/9 plates: | ||
| - | *pSB1K3.TAT⋅SOD⋅His: TAT⋅SH 1-5 | + | *pSB1K3.N-TAT⋅SOD⋅His: TAT⋅SH 1-5 |
| - | *pSB1K3.Tra10⋅SOD⋅His: | + | *pSB1K3.N-Tra10⋅SOD⋅His: Tra10⋅SH 1-5 |
*pEX.SOD 1-4 | *pEX.SOD 1-4 | ||
Revision as of 19:07, 16 September 2010
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Andreas
Assembly of new parts
Gel verification of part digestions
Ran gels of digestion samples in parallel with undigested samples to verify successful digestions and insert sizes.
Gel 1
&4 μl λ; 3 μl sample.
λ = O'GeneRuler 1 kb DNA ladder.
1 % agarose, 110 V
Gel 2
&4 μl λ; 3 μl sample.
λ = O'GeneRuler 1 kb DNA ladder.
1 % agarose, 110 V
Results
Successful digestion with corresponding bands for all digested samples. N-TAT and N-Tra10 not verified, since gel was run too far, but plasmid linearization should indicate successful digestion.
Most samples show somewhat incomplete digestion. For digestions with FastDigest enzymes, this may be an indication of old/inactive enzymes. Especially PstI should be analyzed for activity.
Colony PCR
Picked new colonies for colony PCR from 14/9 plates:
- pSB1K3.N-TAT⋅SOD⋅His: TAT⋅SH 1-5
- pSB1K3.N-Tra10⋅SOD⋅His: Tra10⋅SH 1-5
- pEX.SOD 1-4
Standard colony PCR settings.
- Elongation: 1:30
Gel verification
4 μl λ; 5 μl sample.
1 kb λ = O'GeneRuler 1 kb DNA ladder; 50 bp λ = GeneRuler 50 bp DNA ladder.
4 μl λ; 5 μl sample.
λ = O'GeneRuler 1 kb DNA ladder.
4 μl λ; 5 μl sample.
λ = O'GeneRuler 1 kb DNA ladder.
