Difference between revisions of "C-Fos Methylation"
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| − | ==Rat c-fos Primers== | + | ==Rat c-fos Primers used by Alcala and Houpt== |
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| + | Forward: | ||
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| + | Reverse: | ||
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| + | Product size: | ||
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| + | PCR conditions for use with HotStarTaq mix (progam CFOSGENO on thermocycler) | ||
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| + | 95° for 15 min to activate Taq | ||
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| + | Repeat 35 times: | ||
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| + | :94° for 1 min | ||
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| + | :59° for 1 min | ||
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| + | :72° for 1 min | ||
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| + | :72° for 10 min final extension | ||
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| + | 4° for overnight | ||
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| + | ==Rat c-fos Primers from Literature== | ||
From Shimizu et al., Cancer Sci 2007; 98: 1318–1322 | From Shimizu et al., Cancer Sci 2007; 98: 1318–1322 | ||
Revision as of 17:16, 23 January 2009
DNA Methylation sites in c-Fos promoter
Approximate MW:
38,337 daltons (unmethylated)
38,252 daltons (methylated)
each methylation site causes 15 dalton decrease in MW 
Rat c-fos Primers used by Alcala and Houpt
Forward:
Reverse:
Product size:
PCR conditions for use with HotStarTaq mix (progam CFOSGENO on thermocycler)
95° for 15 min to activate Taq
Repeat 35 times:
- 94° for 1 min
- 59° for 1 min
- 72° for 1 min
- 72° for 10 min final extension
4° for overnight
Rat c-fos Primers from Literature
From Shimizu et al., Cancer Sci 2007; 98: 1318–1322
Bisulfite sequencing (Genomic DNA)
GenBank AF126534
F: 5′-TATTTATAGGTGAAAGTTATAGATTG-3′
R: 5′-CACTAATAAAAACTACAAAACAAAACT-3′
Annealing temperature 54
Product size 292 bases
4-23-08 -- could NOT confirm with blast -- need to check earlier references
RT-PCR (from RNA)
GenBank X06769
F: 5′-TTGCGCAGATCTGTCCGTCT-3′
R: 5′-GTTGATCTGTCTCCGCTTGG-3′
Annealing temperature 65
4-23-08 -- confirmed with blast