SBB11Ntbk-VinidhraMani
--Vinidhra Mani 13:47, 17 February 2011 (EST)
2/17/11 Today, for the completed PCR reactions, the analytical gel was prepared and run. (parts sb1111 and sb1135 for P_oppBCF and P_phrB). 2ul of the PCR product and 5ul of loading dye were added to a 500ul tube, as preparation for analytical gel.
In addition, the digest for the toxic gene (lacZ) was done. 8ul of the PCR product (pBca9145-jtk2791) was mixed with 1ul NEB buffer and digested with 0.5ul each of EcoR1 and BamHI. Subsequently, this was placed in the thermocycler at 37C for 1 hour.
--Vinidhra Mani 14:08, 15 February 2011 (EST)
2/15/11 Today, we did our first simple PCR reactions. I completed the reaction for part sbb1111 and sbb1135, the P_oppBCF and P_phrB promoters, respectively. The PCR protocol listed on our SBB homepage was used. Hopefully this will work the first time! I still have one part to make given that this works, that is the Eco/Bam transfer for the lacZ gene.--Vinidhra Mani 14:07, 15 February 2011 (EST)