|Table of Contents|

Optimization and Establishment of cDNA-AFLP Technical System in Solanum melongena(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2012年19期
Page:
108-112
Research Field:
Publishing date:

Info

Title:
Optimization and Establishment of cDNA-AFLP Technical System in Solanum melongena
Author(s):
WANG Zhong1; 2; YANG Qing2; WANG Ren1; ZHENG Yu-hong1
1.Research Center of Systematic and Evolutionary Botany，Institute of Botany，Chinese Academy of Sciences，Nanjing，Jiangsu 210014；
2.State Key Laboratory of Crop Genetics and Germplasm Enhancement，College of Life Sciences，Nanjing Agricultural University，Nanjing，Jiangsu 210095
Keywords:
Solanum melongena; cDNA-AFLP; system optimization
PACS:
S 641.103.6
DOI:
-
Abstract:
In this study，the key facters affecting cDNA-AFLP on establishing cDNA-AFLP analysis system for roots were analyzed in Solanum melongena with high resistance to Verticillium wilt，and a distinct electrophoretogram of cDNA-AFLP was obtained.The results showed that total RNA extracted by improved Trizol method was highly pure and complete.The double-stranded (ds) cDNA was synthesized using low-cost and efficient replacement and synthesis method.The abundant and repeatable cDNA-AFLP fingerpritings have been obtained when original ligation products were used as template in the preamplification PCR and the preamplification products were diluted to 50 times for selective amplification in 25 μL reaction volume containing 2 μL MgCl2 (25 mM) and 0.2 μL dNTPs (10 mM).

References:

[1]Bachem C W B，Van Derhoeven R S，De Bruijn S M，et al.Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP，analysis of gene expression during potato tuber development [J].Plant Journal，1996,9(5):745-753.

[2]Bachem C W B，Oomen R J F J，Visser R G F.Transcript imaging with cDNA-AFLP：a step-by-step protocol[J].Plant Molecular Biology Reporter,1998,16(2):157-173.

[3]Durrant W E，Rowland O，Piedras P，et al.cDNA-AFLP reveals a strikingoverlap in race-specific resistance and wound response gene expression profiles [J].The Plant Cell,2000,12(6):963-977.

[4]于虹,丁国华.cDNA-AFLP技术及在差异表达基因克隆上的应用[J].中国农学通报,2007,23(12):77-80.

[5]Nazeem P A，Jose Sherin，Sheeba N K，et al.Differential gene expression for bacterial wilt incidence in tomato (Solanum lycopersicum?L.)revealed by cDNA-AFLP analysis [J].Physiological and Molecular Plant Pathology,2011,8(3):1-7.

[6]Wang L，Zhou B R，Wu L L，et al.Differentially expressed genes in Populus simonii×Populus nigra in response to NaCl stress using cDNA-AFLP [J].Plant Science,2011,2(1):796-801.

[7]李怀芳,刘凤权,郭小密.园艺植物病理学[M].北京:中国农业大学出版社,2001.

[8]Halima B，Jacquemin J M，Baudoin J P，et al.Optimization of a reliable，fast，cheap and sensitive silver staining method to detect SSR markers in polyacrylamide gels[J].Biotechnology Agronomy Society and Environment,2006,10(2):77-81.

Memo

Memo:
-
Last Update: 2014-09-09