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Template:Team:UC Berkeley/Notebook/AL notes
From 2008.igem.org
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A. Made the vector pBca1256 | A. Made the vector pBca1256 | ||
::1. PCR-50 ul | ::1. PCR-50 ul | ||
| - | ::::a. 43 ul water, 1 ul dNTP, | + | ::::a. 43 ul water, 1 ul 10mM dNTP, 5 ul 10x Expand Buffer, 1 ul 10uM ca1246F, 1ul 10uM ca1246R, 0.75 ul polymerase, 0.5 ul template (pBca1256) |
::2. Zymo cleanup | ::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. | ::3. Digest vector using 1ul of EcoRI, BamHI, DpnI and 87 DNA/water solution. Jin ran a gel and said it is fine. | ||
| Line 21: | Line 21: | ||
::1. Added 5x more dNTPs, which competes for Mg in buffer with polymerase. | ::1. Added 5x more dNTPs, which competes for Mg in buffer with polymerase. | ||
| - | ::::a. 18.5 ul water, 2.5 ul | + | ::::a. 18.5 ul water, 2.5 ul 10 mM dNTP, 1 ul buffer, 0.5 ul polyermase, 0.5 ul template, 1ul oligo 1, 1 ul oligo 2 |
B. PCR for parts K112300-K112321-Second Try | B. PCR for parts K112300-K112321-Second Try | ||
| + | |||
| + | ::1. 20.375 ul water, 0.5 ul 10 mM dNTP, 1ul buffer, 0.375 ul polyermase, 0.25 ul template, 1ul oligo 1, 1ul oligo 2 | ||
| + | |||
==[[User:Aronlau|Aronlau]] 22:00, 6 June 2008 (UTC)== | ==[[User:Aronlau|Aronlau]] 22:00, 6 June 2008 (UTC)== | ||
Revision as of 16:56, 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 10mM dNTP, 5 ul 10x Expand Buffer, 1 ul 10uM ca1246F, 1ul 10uM 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 10 mM dNTP, 1 ul buffer, 0.5 ul polyermase, 0.5 ul template, 1ul oligo 1, 1 ul oligo 2
B. PCR for parts K112300-K112321-Second Try
- 1. 20.375 ul water, 0.5 ul 10 mM dNTP, 1ul buffer, 0.375 ul polyermase, 0.25 ul template, 1ul oligo 1, 1ul oligo 2
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
