- 1 tube, 2 hours, 3 steps protocol
- Picogram inputs
- Reduce potential bias - few PCR amplification cycles needed
- High sensitivity ChIP-seq - low PCR duplication rate
- Great multiplexing flexibility with 48 barcodes (8 nt) included
|MicroPlex v1||MicroPlex v2|
|PCR cycles (50 pg starting material)||20||16|
|Index sequences||6 nucleotides||8 nucleotides|
|PCR reaction volume||75 µl||50 µl|
Reliable detection of enrichments in ChIP-seq
Figure A. ChIP has been peformed with H3K4me3 antibody, amplification of 17 pg of DNA ChIP'd from 10.000 cells and amplification of 35 pg of DNA ChIP'd from 100.000 cells (control experiment). The IP'd DNA was amplified and transformed into a sequencing-ready preparation for the Illumina plateform with the MicroPlex Library Preparation kit. The library was then analysed on an Illumina® Genome Analyzer. Cluster generation and sequencing were performed according to the manufacturer's instructions.
Figure B. We observed a perfect match between the top 40% of True MicroChIP peaks and the reference dataset. Based on the NIH Encode project criterion, ChIP-seq results are considered reproducible between an original and reproduced dataset if the top 40% of peaks have at least an 80% overlap ratio with the compared dataset.