生物学翻译
泛瑞翻译
原文
Amplification was done in a total volume of 25μl reaction mixture containing 2, 5 μl of template DNA, each reaction mixture contained 10.5 μl of premix. 1 μl for each forward primer and each reverse primer (one PCR program with 5 VFs to detecte). The 5 primer pools, with the 30 primer pairs listed in table 2. the pools were divided as follows: pool 1: PAI (925), papA (717), fimH (508), kpsMT III (392), papEF (326), and ibeA (171); pool 2, fyuA (787), bmaE (507), sfa/focDE (410), iutA (302), papG allele III (internal; 258), and K1 (153); pool 3: hlyA (1177), rfc (788), nfaE (559), papG allele I (internal; 4), kpsMT II (272), and papC (205); pool 4: gafD (952), cvaC (697), cdtB (430), focG (3), traT (290), and papG allele II (internal; 190); and pool 5: papG allele I (flanking; 1190), papG alleles II and III (flanking; 1070), afa/draBC (594), cnf1 (498), sfaS (244), and K5 (159). Reactions were heated to 95°C in an automated thermal cycler (MyGene Series Peltier Thermal Cycler, MG96G) for 12 min to activate the AmpliTaq Gold. This was followed by 25 cycles of denaturation (94°C, 30 s), annealing (63°C, 30 s), and extension (68°C, 3 min) and a final extension (72°C, 10 min). Samples were electrophoresed in 1% agarose gels, then stained with ethidium bromide, disdained with distilled water, and photographed by use of an ultraviolet transilluminator and digital capture system (Gel Documentation and Image Analysis System, ChampGel 3200). The sizes of the amplicons were determined by comparing them with a 2000-bp DNA marker
(Takara Biotechnology, DL2, 000 TM DNA Marker), which was run in multiple lanes on the same gel.
译文
PCR扩增体系为25μl，其中模板DNA2.5 μl，buffer、聚合酶等为10.5 μl。 各引物分别1 μl 引物见表2。引物分为5组： 第1组： PAI (925), papA (717), fimH (508), kpsMT III (392), papEF (326), ibeA (171); 第二组, fyuA (787), bmaE (507), sfa/focDE (410), iutA (302), papG allele III (internal; 258), K1 (153); 第三组: hlyA (1177), rfc (788), nfaE (559), papG allele I (internal; 4), kpsMT II (272), papC (205); 第四组：gafD (952), cvaC (697), cdtB (430), focG (3), traT (290), papG allele II (internal; 190); 第五组：papG allele I (flanking; 1190), papG alleles II，III (flanking; 1070), afa/draBC (594), cnf1 (498), sfaS (244), K5 (159). 反应体系于95°C加热12分钟，以激活AmpliTaq Gold聚合酶。 之后，94°C变性30 s，63°C退火30 s，68°C延伸3分钟，25个循环。最后72°C延伸10分钟。 1%的琼脂凝胶电泳分离PCR产物，EB染色后，单蒸水漂洗。凝胶成像系统（ChampGel 3200）拍照。 根据购自Takara公司的2000-bp DNA marker判断扩增子条带大小。
