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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.
| Product title:
| CGI1 LIBRARY
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| Description:
| Human CpG library supplied as live cells
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| Batch number:
| 96/1
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| Date of production:
| July 1996
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Contents of package:
- (a) 100µl aliquot of CGI1library.
(store at -70°C)
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(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
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| Host Strain: | Methylation tolerant E.coli strain XLI-BLUE MRF'
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| Vector: | pGEM-5Zf(-)
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| Average insert size: | 760 bp
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| Insert excision: | NdeI
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| Growth conditions: | LB + 50µg/ml ampicillin
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| Coverage: | ~ 6 fold
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- 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 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
14th Sept 1998 Version 3.31
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