SBB09Ntbk-Eric Hsieh: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
No edit summary |
||
| Line 12: | Line 12: | ||
===PCR=== | ===PCR=== | ||
<pre> | <pre> | ||
1) Combined the following: 24uL ddH2O, 3.3uL 10x buffer "2", 3.3uL dNTPs (2mM), 1uL FP/RP (10uM), 0.5uL | 1) Combined the following: 24uL ddH2O, 3.3uL 10x buffer "2", 3.3uL dNTPs (2mM), 1uL FP/RP (10uM), 0.5uL of pBca1256-Bca1346 DNA. | ||
2) Added 0.5uL Expand Polymerase "1". | |||
3) Used the C4K55 thermocycler protocol for an expected product ~4kb. | |||
</pre> | </pre> | ||
Revision as of 09:32, 23 February 2009
Eric W. Hsieh 11:22, 23 February 2009 (EST)
On 18 February 2009, received and reconstituted the oligos and performed PCR.
Oligos
1) Performed quick spin on tubes to pellet the DNA powder. 2) Added 288uL ddH2O to 288nmol of oligos to reach a final concentration of 100uM. 3) Took out a 1uL aliquot and diluted ten-fold to 10uM for PCR recipe.
PCR
1) Combined the following: 24uL ddH2O, 3.3uL 10x buffer "2", 3.3uL dNTPs (2mM), 1uL FP/RP (10uM), 0.5uL of pBca1256-Bca1346 DNA. 2) Added 0.5uL Expand Polymerase "1". 3) Used the C4K55 thermocycler protocol for an expected product ~4kb.
Eric W. Hsieh 12:07, 11 February 2009 (EST)
1) Created the construction file for the Ice Nucleation protein repeats.
2) Created this notebook page and added a profile picture.
