SBB10Ntbk-Mesa

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Project

2/17/2010

Received Oligos: KRM001, KRM002, KRM003, KRM004

Diluted all oligos to a 100μM concentration

Made 10μM dilutions of KRM001 and KRM 002

9μl H2O
1μl 100μM oligo

Prepared one wobble and one PCR reaction, KRM16 and KRM27 respectively.

Wobble Reaction (KRM16)
29μl ddH2O
5μl 2
5μl dNTPs
5μl KRM003 (100μM)
5μl KRM004 (100μM)
0.75μl 1

PCR Reaction (KRM27)
24μl ddH2O
3.3μl 2
3.3μl dNTPs
1μl KRM003 (10μM)
1μl KRM004 (10μM)
0.5μl 1
0.5μl Template DNA (pEC22-CA42)

2/24/2010

Performed EcoRI/BamHI Digests on sbb28 and sbb29
After, ran 2 preparative gels

10 microliters digestion product
1 microliters undiluted 6x loading buffer

2/22/2010

Labwork

Last time: basic PCR for sbb28, sbb29
This time: ran 2 analytical gels

3 microliters PCR
7 microliters Loading Buffer

While waiting, ran zymo cleanup on the 2 samples, final volume of 30.6 microliters
Stored in 140L Box A
Feb 22 - zymo cleanup products stored in 140L Box A (note from JennaK)
Next time: digestion

Gel Results

Analytical gel images shown below (ladder breakdown on left, gels on right), loaded and imaged by Dorothy Tulanont



Relevant Lanes:

1. ladder
2. sbb28 - looks to be about 1500 bp
3. sbb29 - looks to be about 1000 bp

Both of these values are close to desired lengths, so PCR worked!