title: GSM4683758: Libhp2; synthetic construct; OTHER
library_construction_protocol: We used phage amplification overnight cultures for NGS sample preparation after boiling the phage to extract ssDNA as templates for PCR reactions. For all samples from both Lib 10AA and Lib hP, a barcoding PCR process was performed using primers listed in Supplemental Figure 21.The thermal PCR profile for Lib 10AA was: 98 °C (20 s), 63 °C (15 s), 72 °C (30 s), 15 cycle. The thermal profile for Lib hP was: 98 °C (30 s), 72 °C (30 s), 15 cycle to prevent heterodimerization due to the 24-residue overlap. The number of cycles was determined empirically to prevent product laddering, assessed by agarose gel electrophoresis. PCR products were gel purified on 1.2 % agarose. Libraries were sequenced on a NextSeq or Highseq4000(Illumina) using single-read 50 or 150 base pair reads. A custom sequencing primer was used (order as shown): ATCGGATCCGCCGCTCTGAAAGTACAGATTCTC (reverse, Tm =67 ◦C, GC% = 52) or CTGCAGTATGTGGCATGAATCTGGTGGTGGT (reverse, Tm =67 ◦C, GC% = 52) for Lib 10AA and GTATGACGTCGCCATCCGCAGTTCGAGAAA (forward, Tm =67 ◦C, GC% = 52) for Lib hp.
attributes.tag: GEO Accession
attributes.value: GSM4683758