rpsL 128 prevention (128 replicate 2) - SRA

SRX1946250: rpsL 128 prevention (128 replicate 2)
1 ILLUMINA (Illumina MiSeq) run: 22,645 spots, 3.4M bases, 1.6Mb downloads

Design: 50 ng of purified E. coli genomic DNA was subjected to 10 cycles of PCR using the respective pair of target-specific primers. After purification with a 1.5X ratio of Ampure XP beads (Agencourt #A63880), 4-8 additional rounds of PCR were performed to add the appropriate Illumina indices for multiplexed sequencing. A final cleanup using a 1.0X ratio of Ampure XP beads was performed prior to qPCR quantification and pooling. In all cases, Phusion DNA Polymerase (NEB #M0530L) was used for the first round of PCR, the KAPA SYBR FAST qPCR Master Mix (KAPA Biosystems #KK4602) was used for the second round (indexing) PCR, and the KAPA SYBR FAST Illumina quantification kit (KAPA Biosystems #KK4824) was used for library quantification. Sequencing was performed on Illumina MiSeq instruments with the v2 chemistry.

Submitted by: Wyss Institute

Study: Precise Cas9 targeting enables genomic mutation prevention

PRJNA329016 • SRP078472 • All experiments • All runs

show Abstracthide Abstract

Introduction of additional mismatches between the gRNA "spacer" and the target "protospacer" can confer single nucleotide specificity to Cas9. This specificity may then be used to engineering a "mutation prevention system", in which Cas9 and a tuned gRNA are used to proactively prevent point mutations from arising within an organism or population of organisms.

Sample:

SAMN05389970 • SRS1562687 • All experiments • All runs

Organism: Escherichia coli str. K-12 substr. MG1655

Library:

Name: rpsl-128-2

Instrument: Illumina MiSeq

Strategy: AMPLICON

Source: GENOMIC

Selection: PCR

Layout: SINGLE

Runs: 1 run, 22,645 spots, 3.4M bases, 1.6Mb

Run	# of Spots	# of Bases	Size	Published
SRR3909578	22,645	3.4M	1.6Mb	2017-09-30

ID:: 2803284

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