| 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' (Geneservice Ltd. no 3558)
5'-AAC GCG TTG GGA GCT CTC CCT TAA-3' (Geneservice Ltd. no 3559)
Additional primer stocks can be obtained through Geneservice Ltd.,
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)
-
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
13thJuly 1999 version 1.12
|
