I am to find out the quantification of circulating tumor DNA and cell free DNA through real time PCR, for that purpose in PCR which gene primers should be included for amplification
Asiyah Bukhari
It depends on type, sub type and stage of the cancer or whether chemotherapy has done for patient or not, whether our patient is taking specific medicine or not...
For example one of the most prevalent mutations in NSCLC (non-small cell lung carcinoma) occurs through a specific SNP in EGFR and increases tyrosine kinase property, And for a patient in the early stage of that cancer, real-time PCR primers can be designed for that specific SNP. if it is positive, the therapy will be started.
Quantitative studies on cell free DNA must be carried out with very high sensitivity and specificity, because there are also many circulating free DNAs released by normal cells. We use TaqMan probes in quantitative studies because we need the most specificity.
We can also use real-time PCR for methylation profile of the patient which can be a very nice tool in predictive approaches.
Asiyah Bukhari
It depends on type, sub type and stage of the cancer or whether chemotherapy has done for patient or not, whether our patient is taking specific medicine or not...
For example one of the most prevalent mutations in NSCLC (non-small cell lung carcinoma) occurs through a specific SNP in EGFR and increases tyrosine kinase property, And for a patient in the early stage of that cancer, real-time PCR primers can be designed for that specific SNP. if it is positive, the therapy will be started.
Quantitative studies on cell free DNA must be carried out with very high sensitivity and specificity, because there are also many circulating free DNAs released by normal cells. We use TaqMan probes in quantitative studies because we need the most specificity.
We can also use real-time PCR for methylation profile of the patient which can be a very nice tool in predictive approaches.
This depend on the individual tumor that you study, their type, stage, location, and so on...
Best
Hi. For me, it is quite difficult with Real Time. First of all in clinic. Timing is essential. Please try to work with a samples (blood) that stored not longer that 1 hour.
On the base on my experience ddPCR is better, more sensitive and reproducible. But more expencive.
- Badges
- Science method