GBT19438.1-2004禽流感病毒通用荧光RT-PCR检测方法国家标准规范.pdf

ICS 11.220 B41 GB/T 19438.12004Method of the real-time RT-PCR for the detection of Avian Influenza Virus 2004-2-15 2004-2-15GB/T 19438.12004 I GB/T1.12000 1 CB GB/T 19438.12004 1 RT-PCR-A AA535FAMR3TAMRA5Q RT-PCR 12 000 r/min 3 000 r/min 2 oC8 oC-20 oC 10 L100 L1 000 L Eppendorf1.5 mL GB/T 19438.12004 2 35 1.0 mL PBS1.5 mL Eppendorf 30 min1.5 mL Eppendorf 2.0 g10 mL PBS4 oC3 000 r/min15 min1.5 mL Eppendorf 2 oC8 oC24 h-70 oC3 RTPCRC 1 GB/T 19438.12004 3 n1.5 mL Eppendorfn 600 L200 L200 L5 s4 oC12 000 r/min15 min 6.2.11.5 mL Eppendorf500 L-20 oC6.2.2500 L 4 oC2 000 r/min15 minEppendorf()600 L 75 4 oC12 000 r/min10 minEppendorf 4 000 r/min10 sEppendorf3 minRNA 11 L DEPCRNA2 000 r/min5 sRNA2 hPCR-70 oC RT-PCRTaq2 000 r/min5 sRT-PCRnn 0.2RT-PCRRT-PCR15 L RT-PCR6.2.7RNA10 L RT-PCR 6.3.2PCRRT-PCR 42 oC/30 min 92 oC/3 min 92 oC/10s45 oC/30s72 oC/1min5 92 oC/10s60 oC/30s40 Ct GB/T 19438.12004 4 Ct Ct30Ct GB/T 19438.12004 5 A 0.2 mol/L NaH2PO4H2O 27.6 g1 000 mL B 0.2 mol/L Na2HPO47H2O 53.6 gNa2HPO412H2O 71.6 gNa2HPO42H2O 35.6 g1 000 mL 0.01 mol/LpH7.2 0.2 mol/L A14 mL 0.2 mol/L B36 mL NaCl 8.5 g 1 000 mL GB/T 19438.12004 6 GB/T 19438.12004 7 GB/T 19438.12004 8