Prior to ordering this library we strongly recommend that you read the following publication:
- Nucleic Acids Res. 2005 May 23;33(9):2952-61.
- CpG Island microarray probe sequences derived from a physical library are representative of CpG Islands
annotated on the human genome. Heisler LE, Torti D, Boutros PC, Watson J, Chan C, Winegarden N, Takahashi M,
Yau P, Huang TH, Farnham PJ, Jurisica I, Woodgett JR, Bremner R, Penn LZ, Der SD.
||Human CpG library supplied as live cells
|Date of production:
Contents of package:
- (a) 100µl aliquot of CGI1library. (store at -70°C)
- (b) 100µl aliquot of PCR primer 3558. (store at -20°C)
- (c) 100µl aliquot of PCR primer 3559. (store at -20°C)
- On Arrival:
Contents should be stored at the above conditions until required.
We suggest that aliquots for freezing are prepared at the same time as first thawing.
Specific product information:
||108 cells/ml (107 cells/100µl) in 25% glycerol + 50µg/ml ampicillin in LB broth
||Methylation tolerant E.coli strain XLI-BLUE MRF'
|Average insert size:
||LB + 50µg/ml ampicillin
||~ 6 fold
- Provided as a 10µM solution
5'-CGG CCG CCT GCA GGT CGA CCT TAA- 3' (Source BioScience LifeSciences no 3558)
5'-AAC GCG TTG GGA GCT CTC CCT TAA- 3' (Source BioScience LifeSciences no 3559)
Additional primer stocks can be obtained through Source BioScience LifeSciences, quoting the above reference numbers.
- Amplification of CpG island fragments:
We have found that the amplification of the CpG island fragments can be achieved in a 20µl reaction volume in the following reaction buffer :
The following 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.
- Quality Control:
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 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