SBB12Ntbk-Annie Joseph: Difference between revisions
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==[[User:Annie Joseph| Annie Joseph]] 12:30, 17 February 2012 (EST)== | ==[[User:Annie Joseph| Annie Joseph]] 12:30, 17 February 2012 (EST)== | ||
We did PCR today! | We did initial PCR/PCA today! | ||
For part sbb1216: | For part sbb1216: | ||
Obtained and made a 10 micro molar solution of oligos ca998/osbb1333R and osbb1216F/g00101 for PCR. | Obtained and made a 10 micro molar solution of oligos ca998/osbb1333R and osbb1216F/g00101 for PCR. | ||
Set up two solutions for PCR: | Set up two solutions for PCR: | ||
This one I call A in construction file: | This one I call A in construction file: | ||
Line 23: | Line 27: | ||
0.5uL pBca9145-jtk2768 | 0.5uL pBca9145-jtk2768 | ||
And... | And... | ||
This one is B: | This one is B: | ||
24uL ddH2O | 24uL ddH2O | ||
3.3uL 10x Expand Buffer "2" | 3.3uL 10x Expand Buffer "2" | ||
Line 35: | Line 44: | ||
For part sbb1205 | For part sbb1205 | ||
I set up a solution based on the PCA protocol on open wet ware. | I set up a solution based on the PCA protocol on open wet ware. | ||
38 uL ddH2O | |||
5 ul 10x expand buffer | |||
5 ul 2mM dNTPs | |||
1 ul oligo mixture (100uM total, mixture of oligos after combination of 100uM stocks) | |||
0.75 ul Expand polymerase | |||
Notes: | |||
n/a |
Revision as of 15:43, 16 February 2012
Annie Joseph 16:23, 13 February 2012 (EST)
Annie Joseph 12:30, 17 February 2012 (EST)
We did initial PCR/PCA today!
For part sbb1216:
Obtained and made a 10 micro molar solution of oligos ca998/osbb1333R and osbb1216F/g00101 for PCR.
Set up two solutions for PCR:
This one I call A in construction file:
24uL ddH2O
3.3uL 10x Expand Buffer "2"
3.3uL dNTPs (2mM in each)
1uL ca998, 10uM
1uL osbb1333R, 10uM
0.5uL Expand polymerase "1"
0.5uL pBca9145-jtk2768
And...
This one is B:
24uL ddH2O 3.3uL 10x Expand Buffer "2" 3.3uL dNTPs (2mM in each) 1uL osbb1216F, 10uM 1uL g00101, 10uM 0.5uL Expand polymerase "1" 0.5uL pBjk2741-jtk3346
For part sbb1205
I set up a solution based on the PCA protocol on open wet ware.
38 uL ddH2O
5 ul 10x expand buffer
5 ul 2mM dNTPs
1 ul oligo mixture (100uM total, mixture of oligos after combination of 100uM stocks)
0.75 ul Expand polymerase
Notes:
n/a