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Team:Cambridge/Lab book/Week 12
From 2012.igem.org
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===Monday (10/09/12)=== | ===Monday (10/09/12)=== | ||
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| + | '''[[Team:Cambridge/Protocols/PCRProtocol|PCR of biobrick vector DNA]]''' | ||
| + | |||
| + | ---- | ||
| + | |||
| + | *Attempted to run PCR of the backbone for the biobricks once more. Settings: Annealing temperature: 56°C, elongation time: 30secs. | ||
| + | |||
| + | *After running on gel, saw that while the -8 vector amplification appears to be producing some correctly sized (~2kb) bands, the other two only seem to be forming many primer dimers. Analysis of the sequence of the prefix and suffix revealed a CG palindrome that appears to be causing self annealing. Inserts are not affected, as they have the palindrome at the 5' end of the primer. | ||
| + | |||
| + | *Will retry at higher temperatures tomorrow. | ||
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| + | ''[[Team:Cambridge/Protocols/Gibsonassembly|Gibson assembly of biobricks]] | ||
| + | |||
| + | ---- | ||
| + | |||
| + | *Attempted Gibson assembly for the -8 magnesium riboswitch biobrick with the backbone fragments produced earlier today and the insert fragments produced yesterday. | ||
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| + | '''[[Team:Cambridge/Protocols/TransformationofE.coli|Transformation of e.coli with Gibson products]] | ||
| + | |||
| + | ---- | ||
| + | |||
| + | *E.coli cells transformed with Gibson products produced earlier today. | ||
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| + | *Transformants plated out on 25μ/ml chloramphenicol plates. | ||
===Tuesday (11/09/12)=== | ===Tuesday (11/09/12)=== | ||
Revision as of 00:36, 17 September 2012
| Week: | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 |
|---|
Contents |
Monday (10/09/12)
- Attempted to run PCR of the backbone for the biobricks once more. Settings: Annealing temperature: 56°C, elongation time: 30secs.
- After running on gel, saw that while the -8 vector amplification appears to be producing some correctly sized (~2kb) bands, the other two only seem to be forming many primer dimers. Analysis of the sequence of the prefix and suffix revealed a CG palindrome that appears to be causing self annealing. Inserts are not affected, as they have the palindrome at the 5' end of the primer.
- Will retry at higher temperatures tomorrow.
- Attempted Gibson assembly for the -8 magnesium riboswitch biobrick with the backbone fragments produced earlier today and the insert fragments produced yesterday.
Transformation of e.coli with Gibson products
- E.coli cells transformed with Gibson products produced earlier today.
- Transformants plated out on 25μ/ml chloramphenicol plates.
Tuesday (11/09/12)
Wednesday (12/09/12)
Thursday (13/09/12)
Friday (14/09/12)
Saturday (15/09/12)
Sunday (16/09/12)
