Hello Everyone,
I am Venkat and am trying to work with cells and tissues from the aorta. I am hoping to study telomere lengths from my samples and I was wondering if there were a reliable way to use qPCR for assaying telomere length.
I am looking at commercially available assays and they seem to use PCR's followed by ELISA's or use northern blots.
Any advice or references to published articles would be greatly appreciated.
Thanks,
Venkat
This link gives you more information regarding your work
https://bmjopen.bmj.com/content/bmjopen/9/Suppl_3/118.full.pdf
This review article would give you enough information about available measurement techniques:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4292845/
You can see several methods for absolute telomere length measurement by qPCR in these articles:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3047434/
https://www.ncbi.nlm.nih.gov/pubmed/18476834
You should be careful when using qPCR - PCR-based techniques (similarly to TRF technique) require high-quality DNA that is not compromised by degradation. In fact the available hybridization techniques are currently considered as a gold standard.
Well known and already proven commercial kits are:
1. TeloTAGGG™ Telomere Length Assay
https://www.sigmaaldrich.com/catalog/product/roche/12209136001?lang=en®ion=BG
2. TRAP Assay
http://www.merckmillipore.com/INTL/en/product/TRAPeze-Telomerase-Detection-Kit,MM_NF-S7700?ReferrerURL=https%3A%2F%2Fwww.google.bg%2F&bd=1
Please download Telomere length measurement. by quantitative real time PCR
We did this recently:
Article The differential spatiotemporal expression pattern of shelte...
In contrast to hybridisation techniques, you need much less starting material for the qPCR.
The best method would be Q PCR due to its high through put,fastness and needs few starting material. i do have also aplan to Telomere length measurement study i saw cawthon and o callaghan studies but still i need more clarification how to do it and how to analyze the data after a PCR. any one have any ideas please let me know it would be really appreciated.
- Badges
- Science method