Method for constructing BAC subclone library

Abstract

This invention provides a kind of improved construction method of BAC subclone base. The processing steps are: extraction and purification of BAC DNA, the fragememtation and end trimming of BAC DNA; the recollection of target fragment and the attachment of vector; the transformation of attachment. For example, the BAC subclone base built by corn is with more than 1500 clones. The empty clone and coliform DNA pollution rate is lower than 5%. The inserted fragment is 3kb in average. This method is easy, cheap, fast and effective.

Description

1. Technical field: The present invention relates to the field of biotechnology, in particular to the preparation of BAC subclones. 2. Background technology: Whole genome sequencing of organisms is of extraordinary significance. Through the analysis of whole genome sequences, a large amount of genetic information can be obtained, which lays the foundation for researches such as functional genomics and comparative genomics. Determining the genome sequence of an organism, identifying the biological function of the sequence, and studying the evolutionary law of an organism have become a hot topic in the field of life science research. Clonal walking method is one of the commonly used sequencing methods. The basic steps include: constructing a large genomic DNA fragment BAC library; using the high-density genetic map and the fingerprint of the BAC clone to locate the BAC clone on the chromosome to construct a whole genome physical map based on the BAC clone; construction of th...

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Problems solved by technology

The technical problem to be solved by the present invention is: the existing methods for constructing BAC subcloning library are complex and cumbersome, expensive, and prone to DNA contamination, which affects the quality of the subcloning library

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