Chicken

Product title: CHICKEN CGI LIBRARY
Description: Chicken CpG library supplied as live cells
Batch number: 96/1
Date of production: July 1996


Contents of package:

  • (a) 100µl aliquot of Chicken 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: NdeI
Average insert size: 670 bp
Growth conditions: LB + 50µg/ml ampicillin
Coverage: ~ 5 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 :

GeneAmp PCR Buffer (10x): Tris-HCL pH 8.3 100mM
(Perkin-Elmer) KCL 500mM
MgCl2 15mM
Gelatin 0.01%


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.

30x cycles of: 94º C 1 min
55º C 1 min
72º C 3 min

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 Chicken CGI library is described in:

CpG islands of chicken are on concentrated microchromosomes
McQueen HA, Fantes J, Cross SH, Clark VH, Archibald Al and Bird AP.
Nature Genetics, 1996 12(3), 321-324