Published November 26, 2018 | Version v1

SNCA targeted gDNA: PacBio raw data

  • 1. Pacific Biosciences
  • 2. Department of Neurology, Duke University

Description

The landscape of SNCA transcripts across synucleinopathies: New insights from long reads sequencing analysis

 

gDNA capture using IDT xGen® Lockdown® Probes and single-molecule sequencing

2µg of each gDNA sample was sheared to 6kb using the Covaris g-TUBE and ligated with barcoded adapters. An equimolar pool of 12-plex barcoded gDNA library (2µg total) was input into the probe based capture with a custom designed SNCA gene panel.

A SMRTBell library was constructed using 626ng of captured and re-amplified gDNA (https://www.pacb.com/wp-content/uploads/Procedure-Checklist-%E2%80%93-Multiplex-Genomic-DNA-Target-Capture-Using-IDT-xGen-Lockdown-Probes.pdf).  A total of 3 SMRT Cells (6 hour movie) were sequenced on the PacBio Sequel platform using 2.0 chemistry.    

 

Barcodes used for gDNA demultiplexing

bc1001: gcagtcgaacatgtagctgactcaggtcacCACATATCAGAGTGCG
bc1002: gcagtcgaacatgtagctgactcaggtcacACACACAGACTGTGAG
bc1003: gcagtcgaacatgtagctgactcaggtcacACACATCTCGTGAGAG
bc1004: gcagtcgaacatgtagctgactcaggtcacCACGCACACACGCGCG
bc1005: gcagtcgaacatgtagctgactcaggtcacCACTCGACTCTCGCGT
bc1006: gcagtcgaacatgtagctgactcaggtcacCATATATATCAGCTGT
bc1007: gcagtcgaacatgtagctgactcaggtcacTCTGTATCTCTATGTG
bc1008: gcagtcgaacatgtagctgactcaggtcacACAGTCGAGCGCTGCG
bc1009: gcagtcgaacatgtagctgactcaggtcacACACACGCGAGACAGA
bc1010: gcagtcgaacatgtagctgactcaggtcacACGCGCTATCTCAGAG
bc1011: gcagtcgaacatgtagctgactcaggtcacCTATACGTATATCTAT
bc1012: gcagtcgaacatgtagctgactcaggtcacACACTAGATCGCGTGT

 

Notes

Three PacBio Sequel 1M cells for targeted gDNA data of SNCA gene in multiplexed (PD, DLB, normal) samples.

Files

m54086_170612_231439.sts.xml

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