Targeted sequencing of post-depletion PAM library: VRKG Cas9 with Spa... - SRA

SRX9386127: Targeted sequencing of post-depletion PAM library: VRKG Cas9 with Spacer 1
1 ILLUMINA (NextSeq 500) run: 9M spots, 694.2M bases, 245.1Mb downloads

Design: A single PCR step (15 cycles) generated dual-indexed amplicons spanning the 8-bp randomized PAM and 20-bp molecular identifier of the randomized PAM library; forward and reverse primers provided the appropriate Illumina indices and adapters needed for direct sequencing on a NextSeq 500. Amplicons were size-selected by gel purification before pooling for a multiplexed, paired-end sequencing run (36 x 36) that captured the entire 8-bp PAM region and part of the 20-bp molecular identifier.

Submitted by: NYU Langone Health

Study: Amplicon sequencing data from PAM depletion experiments

PRJNA672767 • SRP289428 • All experiments • All runs

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Paired-end sequencing of PCR amplicons templated off of a PAM library target plasmid used in E. coli plasmid depletion assays for testing SpCas9 variants programmed with one of two different sgRNA spacers. The PAM library harbored a fully randomized 8-bp PAM region flanked by the two protospacers in appropriate orientations, as well as a distal, fully randomized 20-bp molecular identifier; each read pair captured the complete 8-bp PAM sequence and a non-contiguous region with part of the distal molecular identifier. Amplicons derived from both pre- and post-depletion library samples were sequenced.

Sample:

SAMN16576507 • SRS7606110 • All experiments • All runs

Organism: Escherichia coli

Library:

Name: VRKG-Cas9-Spacer1

Instrument: NextSeq 500

Strategy: AMPLICON

Source: GENOMIC

Selection: PCR

Layout: PAIRED

Runs: 1 run, 9M spots, 694.2M bases, 245.1Mb

Run	# of Spots	# of Bases	Size	Published
SRR12921585	9,015,499	694.2M	245.1Mb	2020-10-28

ID:: 12253776

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