GSM3579596: A-2_control for SubAB; Homo sapiens; OTHER
Genomic DNA from the frozen cells was purified by the conventional phenol-chloroform method. First PCR amplification was carried out with 20 cycles using PrimeStar GXL DNA polymerase (Takara, Otsu, Japan) and the following primers (9 forward primers and 1 reverse primer): (Fw) 1stY1R1s0-8: CTACACGACGCTCTTCCGATCT[-/N/NN/NNN/NNNN/NNNNN/NNNNNN/NNNNNNN/NNNNNNNN]TCTTGTGGAAAGGACGAAACACCG (included 0-8 bp random sequence for increasing library complexity); and (Rv) 1stY2as: GCCACTTTTTCAAGTTGATAACGGACTAG. Second PCR amplification was carried out with 10 cycles using the following primers: (Fw) 2nd P5R1s:  AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT; and (Rv) 2nd P7Y2as: CAAGCAGAAGACGGCATACGAGAT[CC/TT/AA/GG]GCCACTTTTTCAAGTTGATAACGGACTAG (included barcodes for multiplexing of different samples). Amplicon sequencing of sgRNA in GeCKO v2.0 library plasmid
GEO Accession
GSM3579596