||MOUSE CGI LIBRARY
||Mouse CpG library supplied as live cells
|Date of production:
Contents of package:
- (a) 100µl aliquot of Mouse CGI library. (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
||E.coli strain SURE MRF' Vector: pGEM-5Zf(-)
vector sites 326 and 2893
Digest : 2567 bp (vector)
436 bp + insert
|Average insert size:
||LB + 50µg/ml ampicillin
||~ 5 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 20 µlreaction 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.
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 Mouse CGI library is described in:
Cross SH, Lee M, Clark VH, Craig JM, Bird AP and Bickmore WA.
The Chromosmal Distribution of CpG islands in the Mouse:
Evidence for Genome Scrambling in the Rodent Lineage
Genomics 1997 40 454-457