Efficient protocols for CAPS-based mapping in Arabidopsis

被引：14

作者：

Baumbusch, LO

Sundal, IK

Hughes, DW

Galau, GA

Jakobsen, KS

机构：

[1] Univ Oslo, Div Mol Biol, N-0315 Oslo, Norway

[2] Univ Georgia, Dept Bot, Athens, GA 30602 USA

来源：

PLANT MOLECULAR BIOLOGY REPORTER | 2001年 / 19卷 / 02期

关键词：

Arabidopsis; CAPS; DNA extraction; ecotype polymorphisms; gene mapping; PCR conditions;

D O I：

10.1007/BF02772156

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Positional cloning continues to be an essential method for gene identification and characterisation. The introduction of PCR-based techniques such as Amplified Fragment Length Polymorphism (AFLP), Simple Sequence Length Polymorphisms (SSLP) and Cleaved Amplified Polymorphic Sequences (CAPS) has greatly increased the efficiency of gene mapping in arabidopsis. To develop the CAPS marker approach further, we have altered several critical mapping parameters. Efficiency was improved by using a small volume of dry seed for DNA extraction instead of the commonly used vegetative tissue. Reproducibility of PCR reactions was enhanced by faster and reduced protocols for PCR and restriction enzyme digestion and optimisation of PCR conditions for over 50 CAPS primer pairs. Finally, the density of genetic markers was increased by providing polymorphic information for all CAPS markers in arabidopsis ecotypes Wassilewskija (Ws), Columbia (Col) and Cape Verde Islands (Cvi).

引用

页码：137 / 149

页数：13

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