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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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, ?? ul buffer, 1 ul ca1246F, 1ul ca1246R, 0.75 ul polymerase, 0.5 ul template (pBca1256)   
+
::::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 DNTP, 1ul buffer, 0.5 ul polyermase, 0.5 ul template, 1ul oligos
+
::::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.


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