Design: Whole genome sequencing (WGS) libraries were prepared with Illumina-compatible NEXTflex DNA sequencing kit (BIOO Scientific, Austin, Texas, U.S.A.) at the Genotypic Technology Pvt. Ltd., Bangalore, India. Briefly, approx. 1 g of genomic DNA was sheared using Covaris S2 sonicator (Covaris, Woburn, Massachusetts, USA) to generate approx. fragment size distribution from 300 bp to 600 bp. The fragment size distribution was checked on Agilent 2200 TapeStation with D1000 DNA screen tapes and reagents (Agilent Technologies, Palo Alto, CA, USA) and subsequently purified using HighPrep magnetic beads (Magbio Genomics Inc, USA). The purified fragments were end-repaired, adenylated and ligated to Illumina multiplex barcode adaptors as per NEXTFlex DNA sequencing kit protocol.The adapter-ligated DNA was purified with HighPrep beads and then size selected on 2% low melting agarose gel and cleaned using MinElute column (QIAGEN). The resultant fragments were amplified for 10 cycles of PCR using Illumina-compatible primers provided in the NEXTFlex DNA sequencing kit. The final PCR product (sequencing library) was purified with HighPrep beads, followed by library quality control check. The Illumina-compatible sequencing library was initially quantified by Qubit fluorometer (Thermo Fisher Scientific, MA, USA) and its fragment size distribution was analyzed on Agilent TapeStation. Finally, the sequencing library was accurately quantified by quantitative PCR using Kapa Library Quantification Kit (Kapa Biosystems, Wilmington, MA, USA). The qPCR quantified library was subjected to sequencing on an Illumina sequencer for 150 bp paired-end chemistry.
Submitted by: DEPARTMENT OF BIOCHEMISTRY, ALL INDIA INSTITUTE OF MEDICAL SCIENCES(AIIMS), NEW DELHI, INDIA
Study:
Pavo cristatus Genome sequencing and assemblyshow Abstracthide AbstractWe performed a de-novo whole genome sequencing from blood of the Indian blue peacock (Pavo cristatus)
Sample:
Model organism or animal sample from Pavo cristatusLibrary:
Name: SKPea2016_SI
Instrument: Illumina HiSeq 4000
Strategy: WGS
Source: GENOMIC
Selection: PCR
Layout: PAIRED
Runs:
1 run, 489.1M spots, 146.7G bases, 57.6Gb