Kun:LabNotes/MONOD/2013-11-22: Difference between revisions
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===BSPP capture=== | ===BSPP capture=== | ||
*Alan performed the first experiment on five cancer cell lines and three whole blood samples. Note that Alan also called this probe set as GP1: [[AlanFung:LabNotes/Capturing/2013-12-23]]; [[AlanFung:LabNotes/Capturing/2014-1-14]] | *Alan performed the first experiment on five cancer cell lines and three whole blood samples. Note that Alan also called this probe set as GP1: | ||
**[[AlanFung:LabNotes/Capturing/2013-12-23]]; | |||
**[[AlanFung:LabNotes/Capturing/2014-1-14]] | |||
===Data analysis=== | ===Data analysis=== |
Revision as of 03:46, 19 March 2014
MONOD round 1
Target identification
- Data used:
- Cancer data:
- GBM: U87 (ENCODE RRBS Hudson Alpha; ENCODE 450k)
- Pancreatic cancer: PANC1 (ENCODE RRBS UW & Hudson Alpha; ENCODE 450k)
- Whole blood data: GSE30253 RRBS data; GSE31263 WGBS data
- Cancer data:
- DMSs and DMS clusters identification:
- I wrote a simple script that takes the average of all existing data for each CpG site, and reported the sites at which the methylation difference was greater than 0.8 between cancer and whole blood.
./find_DMS_MONOD_v1.pl > MONOD_v1_DMS.txt
- These sites were then grouped into DMS clusters.
./extract_clusters.pl MONOD_v1_DMS.txt > MONOD_v1_DMS_clusters.txt
Probe design
- Dinh went through one round of design using ppDesigner v1.1. However, upon close inspection, I found that some capturing arms contain long stretches of poly-Ts or poly-Gs.
- I modified get_probes.pl, and filtered out any candidate capturing arm that has over 6 A/T/G/Cs.
- I repeated the probe design.
- PANC_GBM_DMS: Target list
- Set I: gap size = 400-450bp, flanking len = 250bp; ppDesigner Output + probe assembly script V6 = Probe sequences: 3,514 probes
- Set II: gap size = 125-175bp, flanking len = 125bp; ppDesigner Output + probe assembly script V4 = Probe sequences: 6,960 probes
- CRC_DMS: Target list.
- Set I: gap size = 400-450bp, flanking len = 300bp; ppDesigner Output + probe assembly script V6 = Probe sequences: 148 probes
- Set II: gap size = 125-175bp, flanking len = 175bp; ppDesigner Output + probe assembly script V4 = Probe sequences: 149 probes
- PANC_GBM_DMS: Target list
- MONOD v1 oligo pool ordered from Custom Array.
BSPP capture
- Alan performed the first experiment on five cancer cell lines and three whole blood samples. Note that Alan also called this probe set as GP1: