|
Status |
Public on Jan 01, 2014 |
Title |
LnCAP SFN-2 |
Sample type |
genomic |
|
|
Channel 1 |
Source name |
MeDIP DNA from LnCAP SFN treatment
|
Organism |
Homo sapiens |
Characteristics |
cell line: LnCAP
|
Treatment protocol |
Cells were treated with 15uM SFN
|
Growth protocol |
LnCAP cells were passaged in RPMI media containing 10% fetal bovine serum
|
Extracted molecule |
genomic DNA |
Extraction protocol |
Genomic DNA was isolated using DNeasy Blood & Tissue Kit (Qiagen), and fragmented using MseI restriction enzyme digestion to yield DNA fragments between 200 to 1,000 bp. Methylated DNA was enriched using methyl-DNA immunoprecipitation (MeDIP) according to manufacturer’s protocol. MeDIP-enriched DNA as well as input DNA was amplified with GenomePlex Complete Whole Genome Amplification kit (Sigma).
|
Label |
Cy5
|
Label protocol |
MeDIP and input DNA were labeled using NimbleGen Dual-Color DNA Labeling Kit per manufacturer's protocol. MeDIP DNA was labeled with Cy5, input DNA was labeled with Cy3
|
|
|
Channel 2 |
Source name |
Input DNA from LnCAP SFN treatment
|
Organism |
Homo sapiens |
Characteristics |
cell line: LnCAP
|
Treatment protocol |
Cells were treated with 15uM SFN
|
Growth protocol |
LnCAP cells were passaged in RPMI media containing 10% fetal bovine serum
|
Extracted molecule |
genomic DNA |
Extraction protocol |
Genomic DNA was isolated using DNeasy Blood & Tissue Kit (Qiagen), and fragmented using MseI restriction enzyme digestion to yield DNA fragments between 200 to 1,000 bp. Methylated DNA was enriched using methyl-DNA immunoprecipitation (MeDIP) according to manufacturer’s protocol. MeDIP-enriched DNA as well as input DNA was amplified with GenomePlex Complete Whole Genome Amplification kit (Sigma).
|
Label |
Cy3
|
Label protocol |
MeDIP and input DNA were labeled using NimbleGen Dual-Color DNA Labeling Kit per manufacturer's protocol. MeDIP DNA was labeled with Cy5, input DNA was labeled with Cy3
|
|
|
|
Hybridization protocol |
Sample hybridization were done using Nimblegen Hybridization System 4 (Roche)
|
Scan protocol |
Array scanning were done using Axon GenePix Pro 4200A (Molecular Device)
|
Description |
MeDIP-chip LnCAP SFN 48h
|
Data processing |
Arrays were processed using Nimblegen's standard default protocol for Nimblescan data extraction. Attached .gff files with scaled, log2 (ChIP/Input) ratio.
|
|
|
Submission date |
May 16, 2013 |
Last update date |
Jan 01, 2014 |
Contact name |
Carmen Wong |
E-mail(s) |
[email protected]
|
Phone |
541-737-4189
|
Organization name |
Oregon State University
|
Department |
Biological and Population Health Sciences
|
Lab |
Emily Ho
|
Street address |
103 Milam Hall
|
City |
Corvallis |
State/province |
OR |
ZIP/Postal code |
97331 |
Country |
USA |
|
|
Platform ID |
GPL17148 |
Series (1) |
GSE47017 |
MeDIP-Chip of PrEC, LnCAP, and PC3 cells treated with vehicle control, sulforaphane, or 3,3’-diindolylmethane |
|