Hi all,
I am facing a problem here and might need TP53 expert's opinion. I will need to amplify the DNA binding domain of TP53 (exon 5 - 9) in just 3 reactions. Based on the different published papers, usually the exon is amplified one by one in single reaction. I heard from my superior that someone in the world could just amplify the exon 1 - 10 of TP53 in just 3 reactions.
Hopefully will get some enlightenment from the TP53 expert.
Tq.
Hi Zx
the human TP53 gene on one PCR...yes it can be largely done since the gene doesn't span a large region in the genome (see attachment). it depends on what you'll do after the PCR but this PCR can be done by LonRangePCR (https://international.neb.com/applications/dna-amplification-pcr-and-qpcr/specialty-pcr/long-range-pcr).
fred
Hi Zx,
ok 8kb is too much for sanger sequencing for which you'll need to cut by 500bp amplicons. Primers have been described in this paper (http://cancerres.aacrjournals.org/content/canres/suppl/2012/01/23/0008-5472.CAN-11-2266.DC1/T2_55K.pdf).
fred
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