SBB10Ntbk-Mesa
From OpenWetWare
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)
2/22/2010
Zymo Clean Up on PCR (KRM27)
Elute in 33μl
Small Fragment Zymo on Wobble (KRM16)
Elute in 50μl
Ran an Analytical Gel for Purified PCR (KRM27) and Wobble (KRM16) samples
3 μl Purified PCR (KRM27) 7 μl Loading Buffer 3 μl Purified Wobble (KRM16) 7 μl Loading Buffer
Purified Samples were stored in140L Box A
Gel Results
KRM27 expected size is 987bp (Second to far right lane) KRM16 expected size is 115bp (Far right lane)
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
