遺伝診断法: |
genome DNA 100ng/μl Control DNA genome DNA 100ng/μl 以前にtyping済のTg genome Plasmid DNA 5ng/μl pBS NSE Q64C Primer F NSE5'NCF 10μM GAGGAGGAGTGGGAATTTTG primer R HAepiR 10μM TTTTTCGGTACCCAGGCTGG reaction mix DNA 1 μl genome 100ng or plasmid 5ng 10 x Ex bufer 2 μl 1x dNTP(25mM) 1.6 μl 200μM Primer F 2 μl 20pmol Primer R 2 μl 20pmol Ex Taq 0.4 μl D.W. 11 μl total 20 μl PCR reaction 95℃-3min. 95℃-1min. 61℃-30sec. 72℃-1min. x 25cycles 72℃-10min. 4℃-Forever E.P. gel: 1.5%Agarose,TAE gel sample: PCR product 5μl + 6x LoadingB 1μl/ lane(25wellの場合) Marker: 100bp DNA Ladder(450ng/lane,25wellの場合) Voltage: 100v time: 25-30min. Target band about 700bp |