|Table of Contents|

A IMPROVED SMART METHOD TO CONSTRUCTIONFULL-LENGTH cDNA LIBRARY FOR LARGE CLONES(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2006年03期
Page:
223-227
Research Field:
Publishing date:

Info

Title:
A IMPROVED SMART METHOD TO CONSTRUCTIONFULL-LENGTH cDNA LIBRARY FOR LARGE CLONES
Author(s):
Dong Zhimin 1;  2 　 Li Yinghui 2 　 Zhang Baoshi 1 　 Guan Rongxia 2 　 Chang Ruzhen 2 　 Qiu Lijuan 2
(1. College of Agronomy , Shenyang Agricultural University , Shenyang 110161 ;2. National Key Facility of Crop Gene Resources and Genetic Improvement ,Key Laboratory of Crop Germplasm &Biotechnology (MOA)/ Institute of CropSciences, Chinese Academy of Agricultural Sciences , Beijing 100081)
Keywords:
Improved SMART ; Amplified cDNA size fractionation; Full-length cDNA library ; Largeclones
PACS:
-
DOI:
10.11861/j.issn.1000-9841.2006.03.0223
Abstract:
It is difficult to obtain large clones among the SMART library due to PCR amplification impai-ring the proportion of large clones. According to this, SMART method was improved based on the primarySMART method. The soybean leaves cDNAs were separated into three parts based on the size distributionof amplified dscDNA by agarose gel size fractionation. Each of three parts was ligated respectively to vec-tor and transformed to Ecoli. The sub-library contain about 1. 0 ×10 6 clones. The recombination ratio wasnearly 99%and full-length ratio was 53. 6 %which was correspondence with that of primary SMART li-brary. The inserts size ranged between 0. 5kb to 3. 5kb and the size of largest inserts was about 1. 5kb lar-ger than that of primary SMART method. The improved SMART method increased obviously the largeclone proportion among the library which was in favor of soybean fuctional genomics research.

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Last Update: 2016-12-15