GBT 19438.1-2004 禽流感病毒通用荧光RT-PCR检测方法

ICS11.220B41GB/T19438.1—2004ÎÌóÐÝÎMethodofthereal-timeRT-PCRforthedetectionofAvianInfluenzaVirus2004-2-152004-2-15GB/T19438.1—2004IGB/T1.1—20001CBGB/T19438.1—20041ÎÌóÐÝÎïÎÌóÐÝÎîRT-PCR-ݬÎÒßÜÛÐÝÐÞÍAÜÒßíAA5’3’5’FAMR3’TAMRA5’QÐÝÎÎÏÎÐÝÎë’í’ÎÎÏÎÏÎììòïRT-PCR12000r/min3000r/min2oC8oC-20oC10µL100µL1000µLEppendorf1.5mLìòîGB/T19438.1—20042ÎÒßÜÛÐÝóîð±ÝÐÞÍïîïî±Ýïë³·²ïððððËñ³ÔéëûÜÛÐÝÙÞêêèîçîóîð±ÝÎÌóÐÝÎïÞëëòïïîïî±Ýïë³·²ïòë³ÔÛ°°»²¼±®ºëòîëòîòï351.0mLPBS1.5mLEppendorfëòîòîëòîòíÛ°°»²¼±®ºëòíëòìëòìòï30min1.5mLEppendorfëòìòî2.0g10mLPBS4oC3000r/min15min1.5mLEppendorfëòë2oC8oC24h-70oC3êêòïRTPCRCêòî1GB/T19438.1—20043êòîòïn1.5mLEppendorfnêòîòî600L200L200L5s4oC12000r/min15minêòîòí6.2.11.5mLEppendorf500L-20oC6.2.2500Lêòîòì4oCï2000r/min15minEppendorf()600L75êòîòë4oC12000r/min10minEppendorfêòîòê4000r/min10sEppendorf3minRNAêòîòé11LDEPCRNA2000r/min5sRNA2hPCR-70oCêòíêòíòïRT-PCRTaq2000r/min5sRT-PCRnnÎÌóÐÝÎïëµÔðòîëµÔ0.2ë²RT-PCRRT-PCR15µLêòíòîRT-PCR6.2.7RNA10µLëðð®ñ³·²íð­êòíòíRT-PCR6.3.2PCRRT-PCR42oC/30min92oC/3min92oC/10s45oC/30s72oC/1min592oC/10s60oC/30s40CtGB/T19438.1—20044ééòïéòîéòîòïCtéòîòîCt28.0éòíéòíòïCtéòíòîCt30éòíòíCt30CtGB/T19438.1—20045ßßòïA0.2mol/LNaH2PO4·H2O27.6g1000mLßòîB0.2mol/LNa2HPO4·7H2O53.6gNa2HPO4·12H2O71.6gNa2HPO4·2H2O35.6g1000mLßòí0.01mol/LpH7.20.2mol/LA14mL0.2mol/LB36mLNaCl8.5g1000mLGB/T19438.1—20046ÞÞòïìèíð³ÔïÜÛÐÝï³ÔïÎÌóÐÝÎéëðÔïÎÌóÐÝÎïñïîïîÔïï³ÔïÎÒßï³ÔïÞòîÞòîòïÎÒßìÞòîòîÜÛÐÝïûÜÛÐÝÎÒßÞòîòíÎÌóÐÝÎÞòíÞòìÞòìòïÞòìòîÎÌóÐÝÎGB/T19438.1—20047ÝÎÌóÐÝÎÝòïÎÒßîëðì¸ÎÒßÝòîÝòîòïîèóîðìïîððð®ñ³·²ðòëÔïðÔëÔîðÔîðÔîððÔîððÔïðððÔøÝòîòîîèóîðíððð®ñ³·²ðòëÔïðÔëÔîðÔîðÔîððÔîððÔïðððÔøÝòîòíÐÝÎÝòíGB/T19438.1—20048Ýòíòïø÷÷øîëì²³÷Ýòíòîøîëì²³÷ÝòíòíÎÌóÐÝÎ