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ERX006574: Cancer Genomics of human
4 ILLUMINA (Illumina Genome Analyzer II) runs: 40.9M spots, 6.5G bases, 4.7Gb downloads

Design: Illumina Paired-end Sequencing of Human Cell Line HCC1954
Submitted by: INCTC
Study: Distinct patterns of somatic alterations between normal and tumor genomes
show Abstracthide Abstract
The main objective of this project is to develop a sequencing strategy to efficiently define the complete catalog of mutations present in the breast tumor cell line HCC1954. Strategies for targeting gene rich regions of the genome, such as exon capture and construction of reduced representation libraries will be combined with shotgun and paired-end sequencing. Parallel sequencing of a lymphoblastoid cell line (HCC1954BL), derived from the same breast cancer patient, will be use to accurately distinguish mutations from polymorphisms.
Sample: Human cell line HCC1954
SAMEA767602 • ERS013009 • All experiments • All runs
Organism: Homo sapiens
Library:
Name: HGWPI
Instrument: Illumina Genome Analyzer II
Strategy: WGS
Source: GENOMIC
Selection: RANDOM
Layout: PAIRED
Construction protocol: A total of 5 μg of genomic DNA from HCC1954 and HCC1954BL cell lines was randomly sheared using a Biorupter according to the manufacturer’s instructions. The fragmented DNA was end-repaired using Klenow and T4 DNA polymerases and phosphorylated at the 5’ end with T4 polynucleotide kinase. A 3’ overhang was created using 3’-5’ exonuclease-deficient Klenow fragment and Illumina paired-end adaptor oligonucleotides were ligated to the created sticky ends. DNA fragments of ~200bp were size selected in 8% polyacrylamide gels and eluted from the gel overnight. Size-selected DNA was PCR amplified for 18 cycles to enrich for adapter-modified DNA fragments. A paired-end flow cell was prepared on the supplied cluster station according to the manufacturer’s protocol. Clusters of PCR colonies were then sequenced on the Illumina GAII sequencing platform using the recommended protocols. Images from the instrument were processed to generate FASTQ sequence files.
Spot descriptor:
forward81  reverse

Runs: 4 runs, 40.9M spots, 6.5G bases, 4.7Gb
Run# of Spots# of BasesSizePublished
ERR0163628,861,7261.4G1Gb2010-12-01
ERR0163639,724,1031.6G1.1Gb2010-12-01
ERR01636410,905,0561.7G1.2Gb2010-12-01
ERR01636511,367,6981.8G1.3Gb2010-12-01

ID:
38038

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