MASA experiment, BRAF mutation. Given the real time pcr curve below from a MASA
ID: 166966 • Letter: M
Question
MASA experiment, BRAF mutation.
Given the real time pcr curve below from a MASA experiment does the sample have a BRAF V600E mutation? Explain your reasoning. What is the 3” nucleotide of the WT and Mutant primer? If the sample had a BRAF V600M mutation would this MASA assay detect it?
Amplification curves 61 2021 222 25 2627 2022 3011 12 Sample ID CP Value V600E MU: MUprimer 23.44 Y600ES ens itivity CTRL 31.17 V600E WT primer No Amp Sample WT primer 25.92 Sample MU primer 25.01 V600E Mu prime W600E se m itivity ctrl Mu prime sample VNT primer sample Mu primer 00E NNT PrimeExplanation / Answer
Real time PCR used to identifying the most common BRAF mutation, V600E, with high sensitivity and specificity of 100%. If V600E is found in the BRAF gene, the BRAF gene mutation test result is positive and thus mutation is takes place in the sample. V600E is one of the most common gene mutations for the BRAF gene and most of the BRAF mutations can be found in human cancers which can encode proteins with elevated kinase activity.
The wild type and mutant primers are as follows:
(5-CCATCGAGATTCTGTAGCTAGACCAAAATC-3)
(5-GCTACAGAATCTCGATGGAGTGGGT-3)
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