High Quality DNA Isolation Method for Chickpea Genotypes
In chickpea breeding genetic studies of individual plants need to be evaluated at the DNA level using molecular markers. A simple and reliable DNA extraction method is a prerequisite. This small-scale method is cetyltrimethylammonium bromide (CTAB)-based and extracts DNA from 1 to 3 folded young leaves processed in a 1.5 ml tube with 0.5 ml of extraction buffer and homogenized using an electric drill. Compared with the micro-prep method the improved mini-prep CTAB method is highly efficient and much cheaper in terms of time, chemical use and labor input. About 49 samples per day can easily be processed by one person. The DNA yield is greater (60 µg per 50-100 µg of fresh leaf tissue) than that obtained from the micro-prep method (50 mg from 5 g of fresh leaf tissue). High quality DNA was obtained and used successfully for restriction endonuclease digestion and polymerase chain reaction amplifications using the mini-prep CTAB method.
Anahtar Kelimeler:
Chickpea, DNA isolation, Molecular markers
High Quality DNA Isolation Method for Chickpea Genotypes
In chickpea breeding genetic studies of individual plants need to be evaluated at the DNA level using molecular markers. A simple and reliable DNA extraction method is a prerequisite. This small-scale method is cetyltrimethylammonium bromide (CTAB)-based and extracts DNA from 1 to 3 folded young leaves processed in a 1.5 ml tube with 0.5 ml of extraction buffer and homogenized using an electric drill. Compared with the micro-prep method the improved mini-prep CTAB method is highly efficient and much cheaper in terms of time, chemical use and labor input. About 49 samples per day can easily be processed by one person. The DNA yield is greater (60 µg per 50-100 µg of fresh leaf tissue) than that obtained from the micro-prep method (50 mg from 5 g of fresh leaf tissue). High quality DNA was obtained and used successfully for restriction endonuclease digestion and polymerase chain reaction amplifications using the mini-prep CTAB method.
Keywords:
Chickpea, DNA isolation, Molecular markers,
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- ISSN: 1300-0152
- Yayın Aralığı: Yılda 6 Sayı
- Yayıncı: TÜBİTAK
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