Life Technologies
BACH2 TaqMan Copy Number Assay
4400291 Hs06142097_cn
I am interested in copy number variations of particular genes in B-cell lymphoma. I have used the Life Technologies Copy Number Assays to validate SNP Array data.
Copy number analysis in lymphoma DNA samples
DNA extracted from fresh frozen lymphoma biopsies
The DNA sample is first diluted to 5ng/ul. To each 384 well of the PCR plate, 5ul of genotyping master mix, 0.5ul of copy number assay, 0.5ul of reference assay and 2ul of nuclease free water is then added to give a total volume of 8ul. 2ul of DNA at 5ng/ul is then added to each well and the plate then run using the following parameters: 95C 10 minutes, (95C 15 seconds then 60C 60 seconds) x40 cycles.
Two reference assays are available and must be validated to choose the optimal reference for each experiment
I obtained good results for BACH2 copy number using the RNase P reference assay, with 2 copies recorded in the calibrator sample and cases of BACH2 gain or loss by SNP array confirmed by RT-PCR.
Rapid validation of copy number variation
Requires some degree of optimisation
Good technique for copy number variation validation, requiring a small amount of DNA sample