SBB12Ntbk-John Seggman

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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.