Employing Digital Droplet PCR to Detect BRAF V600E Mutations in Formalin-fixed Paraffin-embedded Reference Standard Cell Lines.
ddPCR is a highly sensitive PCR method that utilizes a water-oil emulsion system. Using a droplet generator, an extracted nucleic acid sample is partitioned into ~20,000 nano-sized, water-in-oil droplets, and PCR amplification occurs in individual droplets. The ddPCR approach is in identifying sequence mutations, copy number alterations, and select structural rearrangements involving targeted genes. Here, we demonstrate the use of ddPCR as a powerful technique for precisely quantitating rare BRAF V600E mutations in FFPE reference standard cell lines, which is helpful in identifying individuals with cancer. In conclusion, ddPCR technique offers the potential to precisely profile the specific rare mutations in different genes in various types of FFPE samples.
Research Institute of Pharmaceutical Science, Department of Pharmacy, College of Pharmacy, Seoul National University.
Nirmal
N
Rajasekaran
ABION CRO.
Myung Ryurl
MR
Oh
Research Institute of Pharmaceutical Science, Department of Pharmacy, College of Pharmacy, Seoul National University.
Sung-Su
SS
Kim
ABION CRO.
Si Eun
SE
Kim
ABION Inc., R&D Center.
Young Deug
YD
Kim
The Center for Anti-Cancer CDx, N-Bio, Seoul National University.
Hyun-Jeung
HJ
Choi
The Center for Anti-Cancer CDx, N-Bio, Seoul National University.
Bohyun
B
Byun
Research Institute of Pharmaceutical Science, Department of Pharmacy, College of Pharmacy, Seoul National University; The Center for Anti-Cancer CDx, N-Bio, Seoul National University; ykeeshin@snu.ac.kr.
Young Kee
YK
Shin
D002460:Cell Line; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D004252:DNA Mutational Analysis / Q000379:methods*; D005557:Formaldehyde / Q000737:chemistry; D006801:Humans; D009154:Mutation; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics*; D012015:Reference Standards
D005557:Formaldehyde; D004247:DNA; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf