PR8-NS1(1–73)GFP virus sequenced after Covaris shearing - SRA

SRX1142582: PR8-NS1(1–73)GFP virus sequenced after Covaris shearing
1 ILLUMINA (Illumina MiSeq) run: 938,856 spots, 471.3M bases, 307.4Mb downloads

Design: 500 ng of the PR8-NS1(1–73)GFP virus RT-PCR product was sheared with an M220 focused-ultrasonicator (Covaris) set to obtain peak fragment lengths of 300-400 bp. Next, the NEBNext Ultra DNA Library Preparation kit (New England Biolabs) was used to repair the ends and to add Illumina MiSeq-compatible barcode adapters to 100 ng of fragmented DNA. The resulting fragments were size-selected using Agencourt AMPure XP bead sizing (Beckman Coulter). Afterwards, indexes were added in a limited-cycle PCR (10 cycles), followed by purification on Agencourt AMpure XP beads. Fragments were analyzed on a High Sensitivity DNA Chip on the Bioanalyzer (Agilent Technologies) before loading on the sequencing chip. The multiplex sample was heat denatured for 2 min at 96°C before loading on the MiSeq chip. After the 2×250 bp MiSeq paired-end sequencing run, the data were base called and reads with the same barcode were collected and assigned to a sample on the instrument, which generated Illumina FASTQ files (Phred +64 encoding).

Submitted by: DMPR - VIB - UGent

Study: synthetic Influenza A virus Genome sequencing

PRJNA272383 • SRP052023 • All experiments • All runs

show Abstracthide Abstract

Full genome analysis of an influenza A virus expressing GFP from a tri-cistronic NS segment. To reduce the size of this NS segment, a truncated NS1 protein of 73 amino acids combined with a heterologous dimerization domain to increase protein stability was used

Sample: PR8-NS1(1-73)GFP virus

SAMN03281108 • SRS815113 • All experiments • All runs

Organism: synthetic Influenza A virus

Library:

Instrument: Illumina MiSeq

Strategy: AMPLICON

Source: VIRAL RNA

Selection: RT-PCR

Layout: PAIRED

Spot descriptor:

1 forward251 reverse

Runs: 1 run, 938,856 spots, 471.3M bases, 307.4Mb

Run	# of Spots	# of Bases	Size	Published
SRR2155600	938,856	471.3M	307.4Mb	2016-05-25

ID:: 1664869

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