Template:Team:UC Berkeley/Notebook/AL notes

From 2008.igem.org

(Difference between revisions)
(Aronlau 22:08, 11 June 2008 (UTC))
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6/9/2008
6/9/2008
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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
 
 +
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.


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