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Template:Team:UC Berkeley/Notebook/AL notes
From 2008.igem.org
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6/9/2008 | 6/9/2008 | ||
| - | A. Filled out PCR table for parts K112300-K112321 | + | A. Filled out PCR table for parts K112300-K112321 to speed up PCR time when oligos arrive. |
| - | |||
| + | 6/10/2008 | ||
| + | Note: Oligos came and diluted | ||
A. Made the vector pBca1256 | A. Made the vector pBca1256 | ||
| - | ::1. PCR | + | ::1. PCR-50 ul |
| + | ::::a. 43 ul water, 1 ul dNTP, ?? ul buffer, 1 ul ca1246F, 1ul ca1246R, 0.75 ul polymerase, 0.5 ul template (pBca1256) | ||
::2. Zymo cleanup | ::2. Zymo cleanup | ||
| - | ::3. Digest vector using EcoRI, BamHI, DpnI. | + | ::3. Digest vector using 1ul of EcoRI, BamHI, DpnI and 87 DNA/water solution. Jin ran a gel and said it is fine. |
| + | |||
6/11/2008 | 6/11/2008 | ||
| - | A. PCR for parts K112300-K112321 | + | A. PCR for parts K112300-K112321-First Try **Refer to construction files for oligos** |
| + | ::1. Added 5x more dNTPs, which competes for Mg in buffer with polymerase. | ||
| + | |||
| + | ::::a. 18.5 ul water, 2.5 ul DNTP, 1ul buffer, 0.5 ul polyermase, 0.5 ul template, 1ul oligos | ||
| + | |||
| + | B. PCR for parts K112300-K112321-Second Try | ||
==[[User:Aronlau|Aronlau]] 22:00, 6 June 2008 (UTC)== | ==[[User:Aronlau|Aronlau]] 22:00, 6 June 2008 (UTC)== | ||
Revision as of 06:36, 12 June 2008
Aronlau 22:08, 11 June 2008 (UTC)
6/9/2008
A. Filled out PCR table for parts K112300-K112321 to speed up PCR time when oligos arrive.
6/10/2008
Note: Oligos came and diluted
A. Made the vector pBca1256
- 1. PCR-50 ul
- a. 43 ul water, 1 ul dNTP, ?? ul buffer, 1 ul ca1246F, 1ul ca1246R, 0.75 ul polymerase, 0.5 ul template (pBca1256)
- 2. Zymo cleanup
- 3. Digest vector using 1ul of EcoRI, BamHI, DpnI and 87 DNA/water solution. Jin ran a gel and said it is fine.
- 1. PCR-50 ul
6/11/2008
A. PCR for parts K112300-K112321-First Try **Refer to construction files for oligos**
- 1. Added 5x more dNTPs, which competes for Mg in buffer with polymerase.
- a. 18.5 ul water, 2.5 ul DNTP, 1ul buffer, 0.5 ul polyermase, 0.5 ul template, 1ul oligos
B. PCR for parts K112300-K112321-Second Try
Aronlau 22:00, 6 June 2008 (UTC)
6/2/2008 to 6/5/2008
A. Safety Training
B. Cloning Training-PCR, Purification, Digestion, Miniprep, Transformation, Autoclaving
6/4/2008 to 6/6/2008 - Design Oligos/Make Construction file
A. Oligos al0001 to al0024 for parts K112300-K112321
