A BAC end sequencing method

A BAC end sequencing method is disclosed. The method includes following steps of 1) connecting a target segment to a cloning vector to obtain a recombinant vector, 2) subjecting the recombinant vector to digestion with a restriction enzyme, recovering linear DNA segments longer than the cloning vect...

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Hauptverfasser: YANG XIAOHUI, PANG WANFU, YANG YU, JIN LIPING, XU JIANFEI, MA WEIQING, LIU JIE, BIAN CHUNSONG, DUAN SHAOGUANG, GUO XIAO, LI GUANGCUN
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creator YANG XIAOHUI
PANG WANFU
YANG YU
JIN LIPING
XU JIANFEI
MA WEIQING
LIU JIE
BIAN CHUNSONG
DUAN SHAOGUANG
GUO XIAO
LI GUANGCUN
description A BAC end sequencing method is disclosed. The method includes following steps of 1) connecting a target segment to a cloning vector to obtain a recombinant vector, 2) subjecting the recombinant vector to digestion with a restriction enzyme, recovering linear DNA segments longer than the cloning vector to obtain a DNA segment containing two ends of the target segment, wherein the target segment at least comprises two digestion recognition sites of the restriction enzyme and the cloning vector does not have digestion recognition sites of the restriction enzyme, and 3) connecting the DNA segment containing two ends of the target segment to achieve cyclization so as to obtain a plasmid containing the ends of the target segment, and sequencing the plasmid, thus achieving end sequencing of the target segment. Experiments show that BAC end sequencing by the method is 100% successful, and therefore the problem that the success rate of BAC end sequencing is low is overcome, BAC end sequencing is made simple and feasible and is low in cost, and the method is particularly suitable for large-scale BAC end sequencing.
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subjects BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS OR TEST PAPERS THEREFOR
CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES
ENZYMOLOGY
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS
METALLURGY
MICROBIOLOGY
MUTATION OR GENETIC ENGINEERING
PROCESSES OF PREPARING SUCH COMPOSITIONS
SPIRITS
VINEGAR
WINE
title A BAC end sequencing method
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