Methylation tolerant E.coli strain XLI-BLUE MRF'
Average insert size:
Circle grow + 75µg/ml ampicillin
Provided as a 10µM solution 5'-CGG CCG CCT GCA GGT CGA CCT TAA- 3' (Source BioScience UK Ltd. no 3558) 5'-AAC GCG TTG GGA GCT CTC CCT TAA- 3' (Source BioScience UK Ltd. no 3559) Additional primer stocks can be obtained through Source BioScience UK Ltd, quoting the above reference numbers.
We have found that the amplification of the CpG island fragments can be achieved in a 20µl reaction volume in the following reaction buffer : PCR cycle has been successfully used for amplification of the CpG island fragments. We strongly recommend that the conditions are optimized, to allow for lab to lab variation
Our quality control work showed that the above primers with the conditions stated amplified the CpG island fragments successfully. Alternative primers, T7 and SP6 also carry out successful amplification of inserts.
5' -TAA TAC GAC TCA CTA TAG GG- 3' (T7) 5' -GAT TTA GGT GAC ACT ATA G- 3' (SP6)
The database can be found here (link to the attached document)
The construction of the CGI1 library is described in:
Purification of CpG islands using a methylated DNA binding column Cross SH, Charlton JA, Nan X and Bird AP. Nature Genetics, 1994 6 (3), 236-244