We want to detect low abundance of mutations and differentiated expression of genes in tissue samples. We suppose that some mutations and differentiated expression of genes in the cancer cells, but not in the stromal cells. Whole sample DNA or RNA extrations may lead to artificial results, some times false negative and some times false positive. We try laser micro-dissection of the fresh samples to dissect the cancer tissue, but due to the low concentration of the DNA or RNA after extration, analysis can not be conducted.
Hello
See links here , Hope help you
Good Luck
http://link.springer.com/article/10.1007/s11010-012-1282-2
https://asuragen.com/wp-content/uploads/2016/05/A-Unified-and-Streamlined_Latham_AACR16.pdf
http://qpcrupdate.com/bustin-2000.pdf
Huiyi Feng
We have new methods for ultra sensitive qPCR of single base mutants. See website: www.genetagtech.com
If you have access to patient samples we need collaborators to validate our enhanced "liquid biopsy" assays for the major diagnostic mutants related to lung, brain and colon cancer. With these methods, we can reliably detect mutant frequencies as low as 0.01 %.
Let us know if interested.
David Shafer, founder, GeneTAG Technology, Inc. Atlanta, GA USA. [email protected]
Hi Huiyi,
the key point in your study will be the amount of cells you're starting with. since 1ng of DNA means around 150 cells, no need to get a deep sequencing if you just have poor quantities of material.
fred
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