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METHOD FOR ANALYZING STRUCTURE OF MICROBIAL COMMUNITY USING T-RFLP

机译：T-RFLP的微生物群落结构分析方法

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PROBLEM TO BE SOLVED: To provide a T-RFLP (terminal restriction fragment length polymorphism) method in which the community structure of an environmental microorganism can inexpensively be analyzed in a short time.;SOLUTION: The T-RFLP method comprises the following steps. a collecting step S1 of collecting a sample, an amplifying step S3 of amplifying a 16S-rRNA gene with a primer using a fluorescent label of a certain color on the 5'-terminal side and a primer using a fluorescent label of a color different from that of the fluorescent label on the 3'-terminal side, a cleaving step S4 of cleaving at least two sites of a sequence sandwiched between the 5'-terminal side and the 3'-terminal side of the amplified product with a restriction enzyme, a measuring step S5 of measuring the length of the cleaved sequence with a fluorescent DNA sequencer and a microbial species identifying step S6 of determining (identifying) the species or genus of the microorganism in the sample on the basis of the cleaved fragment size on the 5'-terminal side and the cleaved fragment size of the sequence on the 3'-terminal side measured in the measuring step.;COPYRIGHT: (C)2006,JPO&NCIPI
机译：解决的问题：提供一种T-RFLP（末端限制性片段长度多态性）方法，可以在短时间内廉价地分析环境微生物的群落结构。;解决方案：T-RFLP方法包括以下步骤。收集样品的收集步骤S1，扩增步骤S3，其使用在5'-末端侧使用某种颜色的荧光标记物用引物扩增16S-rRNA基因，并使用使用与不同颜色的荧光标记物进行引物扩增的引物在3'末端侧的荧光标记的末端，进行切割步骤S4，该步骤用限制酶切割夹在扩增产物的5'-末端侧和3'-末端侧之间的序列的至少两个位点，测量步骤S5，其利用荧光DNA测序仪和微生物种类测量切割序列的长度。鉴定步骤S6，基于5的切割片段大小，确定（鉴定）样品中微生物的种类或属。 '末端和在测量步骤中测量的3'末端的序列的切割片段大小。;版权所有：（C）2006，JPO＆NCIPI

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