变性高效液相色谱在大通量筛查基因突变中的应用-环境化学.PDF

25 3 Vol. 25, No. 3 2006 5 ENV IRONMENTAL CHEM ISTRY M ay 2006 * 1 1 1 1* * 2 顾 颖 李 延 陈蔚丰 尹大强 赵庆顺 ( 1 , , , 210093; 2 , , , 210061) ( DHPLC) 7p 53 PCR , , , DH PLC. 60 , , 95% . , , p 53, . ( DS) ( SSCP) ( DGGE) . , , . ( DHPLC) [ 1, 2] . , SSCP DNA , DHPLC, , [ 3, 4] . 7p 53 PCR , DHPLCp 53, DHPLC. 1 p53 7 PCR, 173bp, 3, W t, M 1M2, 49bp, 66bp 145bp. 3p 53 7, 5m l LB - 1 ( 100gm l ) , 37 ; 12000g 2m i , ; 40l 1 STE, 40l ( 11) ; 12000g 2m i , PCR . : 5GTTTAACAGTCACATTTTCCT3 (), 5ACAAGAGGAGGAATCAAATA 3 (). PCR : 95 1m i , 94 40s, 52 30s, 72 30s, 30 , 72 10m i . 1l PCR 1%. PCR PCR 95 5m i, 945 20s, 20s 05, 25, DNA . PCR DHPLC, . 5,l DNASep. (T = 5925 ) , T 2 , 59, 595 , 60 605 . m m - 1 - 1 A ( 01mol l ) B ( 01 mol l 25% ) , , 09m lm i - 1 DNASep DNA , 260 m , , , DH PLC, . 2 DHPLCp 53 7 1WAVEMAKER40 173bp 59 , 595, 60 , 605, 615 625 . 50% 99% , * ( . * * : yi dq@ jueduc 3 : 337 [ 1] DNA , 49bp, 66bp 145bp, 59, 595 , 60 , 605. 1 F ig1 M elti g profiles of the fragm e t u der d iffere t temperature M 1M2 PCR W tPCRDNA 11, DHPLC . 2( a, b). 2( a) , , , M 1, , , A= T, G C 3, , . 2( b) ,