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文献和实验Hepes. Harvest cells by putting 20mls of -Ca-Mg PBS, buffered with 0.02M Hepes, and 1mM EDTA on each plate for 10min in the 37 C incubator. After the 10min, collect some of the PBS in a 50ml conical tube before pipeting
Protocols for ET recombination
synthesis, the EMBL oligo service prefers that the 5' most nucleotide is a T or C. Consequently most of our oligos start with a T or C however we do not think this is important for ET cloning efficiencies). (2)The 3' end (the PCR primer) - choose about 18
Protocols for ET recombination
the cells at 2.3 kV (Bio-Rad Gene Pulser), 25 uF with Pulse controller set to 200 ohms) Add 1 ml of LB medium and transfer back into the eppendorf tube. Incubate at 37℃ for 1 to 1.5 hours with shaking. Plate 100ul on suitable antibiot℃ plates, spin
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