Format

Send to:

Choose Destination

SRX2251664: IB_after_3w
1 ILLUMINA (Illumina MiSeq) run: 1.2M spots, 705.7M bases, 404.4Mb downloads

Design: The blood samples collected in the influenza vaccination study by Laserson et al (PMID:24639495) were re-sequenced using the Illumina MiSeq platform. RNA was reverse-transcribed into cDNA using a biotinylated oligo dT primer. An adaptor sequence was added to the 3' end of all cDNA, which contains the Illumina P7 universal priming site and a 17-nucleotide unique molecular identifier (UMI). Products were purified using streptavidin-coated magnetic beads followed by a primary PCR reaction using a pool of primers targeting the IGHA, IGHD, IGHE, IGHG, IGHM, IGKC and IGLC regions, as well as a sample-indexed Illumina P7C7 primer. The immunoglobulin-specific primers contained tails corresponding to the Illumina P5 sequence. PCR products were then purified using AMPure XP beads. A secondary PCR was then performed to add the Illumina C5 clustering sequence to the end of the molecule containing the constant region. The number of secondary PCR cycles was tailored to each sample to avoid entering plateau phase, as judged by a prior quantitative PCR analysis. Final products were purified, quantified with Agilent Tapestation and pooled in equimolar proportions, followed by high-throughput paired-end sequencing on the Illumina MiSeq platform. For sequencing, the Illumina 600 cycle kit was used with the modifications that 325 cycles was used for read 1, 6 cycles for the index reads, 300 cycles for read 2 and a 10%PhiX spike-in to increase sequence diversity.
Submitted by: Yale University
Study: Dynamics of the human antibody repertoire after influenza vaccination
show Abstracthide Abstract
To characterize the dynamics of the immune response to vaccination inhuman volunteers, three individuals were given influenza vaccination and blood samples were taken at -8days, -2d, -1hour, +1h, +1d, +3d, +7d, +14d, +21d, +28d. The samples were sequenced using the Illumina MiSeq platform.
Sample: IB_after_3w
SAMN05924301 • SRS1751537 • All experiments • All runs
Organism: Homo sapiens
Library:
Name: IB_after_3w
Instrument: Illumina MiSeq
Strategy: AMPLICON
Source: TRANSCRIPTOMIC
Selection: PCR
Layout: PAIRED
Runs: 1 run, 1.2M spots, 705.7M bases, 404.4Mb
Run# of Spots# of BasesSizePublished
SRR44317701,163,343705.7M404.4Mb2017-01-24

ID:
3305409

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Support Center