SBB12Ntbk-John Seggman: Difference between revisions
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Prepared the following PCR reaction for SBB1201: | Prepared the following PCR reaction for SBB1201: | ||
24uL ddH2O | 24uL ddH2O | ||
3.3uL 10x Expand Buffer "2" | 3.3uL 10x Expand Buffer "2" | ||
3.3uL dNTPs (2mM in each) | 3.3uL dNTPs (2mM in each) | ||
1uL Oligo 1, 10uM | 1uL Oligo 1, 10uM | ||
1uL Oligo 2, 10uM | 1uL Oligo 2, 10uM | ||
0.5uL Expand polymerase "1" | 0.5uL Expand polymerase "1" | ||
0.5uL Template DNA | 0.5uL Template DNA | ||
Discarded the remainder of the diluted oligos araR-F and araR-R per the "Cloning by PCR" protocol. | Discarded the remainder of the diluted oligos araR-F and araR-R per the "Cloning by PCR" protocol. | ||
Revision as of 12:37, 16 February 2012
John Seggman 14:33, 16 February 2012 (EST)
Made 100uM stocks of oligos obb1226F and obb1226R
Prepared the following wobble reaction for SBB1226:
29 uL water 5 uL Expand 10x Buffer 2 5 uL 10x dNTPs (2 mM in each; 0.2 mM final conc) 5 uL Oligo 1 (100uM) 5 uL Oligo 2 (100uM) 0.75 uL Expand Polymerase 1
Made 100uM stocks of oligos araR-F and araR-R For oligos araR-F and araR-R, made an oligo dilution of:
9uL Water 1uL 100uM oligo
Prepared the following PCR reaction for SBB1201:
24uL ddH2O 3.3uL 10x Expand Buffer "2" 3.3uL dNTPs (2mM in each) 1uL Oligo 1, 10uM 1uL Oligo 2, 10uM 0.5uL Expand polymerase "1" 0.5uL Template DNA
Discarded the remainder of the diluted oligos araR-F and araR-R per the "Cloning by PCR" protocol.
Followed all protocols exactly.
John Seggman 02:23, 15 February 2012 (EST)
This is my next entry.
