Application
ChIP
Detection step
Real-time PCR
Sample
Zebrafish Cell lysate - nuclear (whole embryo shield stage, 6 hours p.f.)
Specification
whole embryo shield stage, 6 hours p.f.
Negative control
We used PCR for zgc15 and igsf genomic regions (see additional information for primer sequences) as a reference control. No H3K4 mono- and di- methylation was expected in these regions based on previously published microarray data
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde 1% in PBST
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde 1% in PBST
Positive control
ChIP sample was 22 times enriched over input control in hoxb3a and 8 times in tiparp genomic region (see additional information for primer sequences).
10-20x enrichment in H3K4 mono- and di- methylation was expected in these regions based on previously published microarray data.
Other product details
Incubation time
12 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: ChIP dilution buffer (Upstate)
Additional data
Additional Notes
Sequences of forward and reverse primers used for assay, 5'to 3':
tiparp-2, H3me1/2 enriched GGAACCGTTGCCTTAAATCTT CACTTAAGCTCTAACGACCCC
hoxb3a-1, H3me1/2 enriched AGTGTGGCGTCAGTAACAGC TCTACAGTTTGCGAGGCTCC
zgc15-2, control ATCAGCCGGTTACCATATAGAAA GCGCCTAGAATACATTTTGCTAT
igsf2-1, control TCTTCGCTGGACTCAAACTG ACAGCTTTCCATCAGTCTTTCA
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
MRS. Daria Onichtchouk
Verified customer
提交于 Apr 01 2015