Product title: PIG CGI LIBRARY
Description: Pig CpG library supplied as live cells
Batch number: 97/1
Date of production: February 1997


Contents of package:

  • (a) 100µl aliquot of Pig 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:

Titre: 108 cells/ml (107 cells/100µl) in 25% glycerol + 50µg/ml ampicillin in LB broth
Host Strain: E.coli strain SURE MRF'
Vector: pGEM-5Zf(-)
Insert excision: Pvu II
vector sites 326 and 2893
Digest = 2567 bp (vector)
436 bp + insert
Average insert size: 500 bp
Growth conditions: LB + 50µg/ml ampicillin
Coverage: ~ 10 fold
Primers:
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 :
Pig data sheet1

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.
Pig data sheet2

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)

 
References:

The construction of the Pig CGI library is described in: McQueen HA, Clark V, Bird A, Yerle M and Archibald A. CpG Islands of the Pig. 1997 Genome Research 7: 924-931