Mini-prep method suitable for a plant breeding program

被引:89
作者
Haymes, KM [1 ]
机构
[1] CTR PLANT BREEDING & REPROD RES,CPRO,DLO,DEPT VEGETABLES & FRUIT CROPS,NL-6700 AA WAGENINGEN,NETHERLANDS
关键词
DNA isolation; PCR; Southern blots;
D O I
10.1007/BF02671664
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Plant breeders need a nondestructive and inexpensive protocol to screen large numbers of plants early after sowing. Isolation of DNA represents one of the limiting steps in this process: normally a person is capable of isolating 25 to 50 samples per day. To speed up the isolation of DNA in marker-assisted plant breeding programs, a CTAB mini-prep was modified into an inexpensive and nondestructive procedure. With this protocol a single individual can isolate 200 to 250 samples per day of high-quality DNA that is immediately suitable for testing by PCR. The amounts of DNA isolated are sufficient fur about 150 PCR amplifications. These improvements are achieved by eliminating time-consuming and noncritical steps in the standard protocol, such as extensive grinding, washing with ethanol, and treatment with RNAse A.
引用
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页码:280 / 284
页数:5
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