1、 ICS 11.220 B41 GB/T 19438.42004H9RT-PCRMethod of the real-time RT-PCR for the detection of Avian Influenza virus subtype H9 2004-2-15 2004-2-15GB/T19438.42004 I GB/T1.12000 1 GB/T19438.42004 1 H9RT-PCR12 3 4H9RT-PCRTaqManH9535FAMR3TAMRA5Q 5 5.1 RNA GB/T19438.42004 2-20 oC PBS1212 15 min10 000 U/mL
2、75%:RNA-20 oC H9RT-PCR1:A 5.2 RT-PCR 12 000 r/min 2 000 r/min (2 oC8 oC-20 oC)10 L100 L1 000 L Eppendorf1.5 mL 6 6.1 6.2 6.2.1 35 1.0 mL PBS1.5 mL Eppendorf 6.2.2 6.2.3 6.3 6.4 6.4.1 30 min1.5 mL Eppendorf 6.4.2 2.0 g10 mL PBS4 oC3 000 r/min15 min1.5 mL Eppendorf 6.5 1 GB/T19438.42004 3 2 oC8 oC24 h
3、-70 oC3 7 7.1 7.2 7.2.1 n1.5 mL Eppendorfn7.2.2 600 L200 L200 L5 s4 oC12 000 r/min15 min7.2.3 6.2.11.5 mL Eppendorf500 L-20 oC6.2.2500 L7.2.4 4 oC2 000 r/min15 minEppendorf()600 L 757.2.5 4 oC12 000 r/min10 minEppendorf7.2.6 4 000 g10 sEppendorf3 minRNA7.2.7 11 L DEPCRNA2 000 r/min5 sRNA2 hPCR-70 oC
4、 7.3 7.3.1 RT-PCRTaq2 000 r/min5 sRT-PCRnn RT-PCR Taq 15 L 0.25 L 41RT-PCRRT-PCR15 L 7.3.2 RT-PCR7.1.7RNA10 L 7.3.3 RT-PCR GB/T19438.42004 4 7.2.2PCRRT-PCR 42 oC/30 min 92 oC/3 min 92 oC/10s45 oC/30s72 oC/1min5 92 oC/10s60 oC/30s40 Ct 8 8.1 8.2 8.2.1 Ct 8.2.2 Ct30 GB/T19438.42004 5 A H9RT-PCR A.1 H9 A.2 A.2.1 A.2.2 A.2.3 A.3 A.3.1 A.3.2 A.3.3
