식품의약품안전청 _KJYDGP DNA Methylation by EZ DNA Methylateion-Gold Kit (ZYMO RESEARCH) 1-2. The Sample list 1-1. The Method of Methylation Control DNA_F1, Treated DNA_F1 두 개의 샘플에 대해서 EZ DNA Methylateion-Gold Kit (ZYMO RESEARCH) 을 사용하여 methylation 시킨 뒤 PCR 실험을 수행하였고, Negative control 로 methylation 처리를 하지 않은 원상태의 Control DNA_F1, Treated DNA_F1 또한 PCR 실험을 수행하였다 Sample Name 1Control DNA_F1-Mehtylated 2Treated DNA_F1-Mehtylated 3Control DNA_F1-unmehtylated 4Treated DNA_F1-unmehtylated
식품의약품안전청 _KJYDGP Primer list 1. DNA Methylation by EZ DNA Methylateion-Gold Kit (ZYMO RESEARCH)
The Result of Secondary PCR 2-1. PCR reaction combination 식품의약품안전청 _KJYDGP PCR Condition Temp.TimeCycles Final Reaction vol. 95 ℃ 5 min 1 30ul Ex-Taq 94 ℃ 30 sec ℃ 30 sec 72 ℃ 1 min 72 ℃ 5 min 1 4℃4℃ Storage 1 PCR 수행 시 Control DNA _F1-Methylated, Treated DNA_F1-Methylated, Control DNA_F1-unmethylated, Treated DNA_F1-unmethylated 4 개의 DNA 샘플과 17 개의 primer set 을 가지고 primary PCR 수행 후 여기서 얻은 PCR product DNA 를 template 로 하여 secondary PCR 을 수행하여 결과를 얻었다
PCR Result 2% 0..5X TAE gel 식품의약품안전청 _KJYDGP The Result of Secondary PCR
The Result of PCR 식품의약품안전청 _KJYDGP kb Ladder 2-4. PCR Result 400bp 300bp 800bp 2% 0..5X TAE gel