Hi everyone. I am seeking to validate the effect on splicing of a TP53 somatic mutation identified with NGS. I am going to sequence the cDNA derived from tissue specimen and I was wandering to use the cDNA extracted from lymphocytes as wild type control (I thought of using lymphocytes because they are easy to recover). Nevertheless, since I have never worked with RNA, I wanted to know if lymphocytes-derived cDNA could be suitable for this purpose.
Many thanks for your help!
Chiara
Hello,
DNA from normal lymphocyte should be a good control to get wt TP53 cDNA. Nevertheless, if you wanted to be very strict, you should use normal DNA from the same tissue type as the original tumor. On the other end, RNA sequencing from the tumor should give you directly some indications as the normal splicing of TP53 is already well established and finding new splice variants should be a good indication that splicing is impaired.
Do not forget that many splice variants have already been described for TP53. It is possible that your mutation has already been tested. If you want to check, contact me by email.
Best
Thierry
Thierry,
Would p53 isoforms be easily detectable using RNAseq or the result might vary depending on the tissue?
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