家畜・家禽のゲノムDNA抽出におけるガラスろ紙法の検討

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In this study, to develop a cost-effective and rapid genomic DNA purification method in animals and birds, we examined the DNA purification method using glass fiber filter proven in plant genomics. Four or eight blood cell samples from each of cattle and chickens were used for the purification. The purification method was basically the same as that described previously (Muramoto et al. 2005). Several points were tested in this method for recovering DNA efficiently, revealing the following: 1)One μg of genomic DNA was purified from the chicken blood cell samples diluted 50-200 times. It could be used for -5kb PCR amplification. Genomic DNA from the chicken blood cell samples diluted 1-10 times could not be used for PCR amplification. In cattle, genomic DNA was purified from the blood cell samples diluted 1-200 times. It could be use for 0.8kb PCR amplification. 2)Concerning concentration of sodium sulfite in sample storage, 10% was acceptable. 3)Drying the filter with the blood cells before purifying improved DNA yield. 4)Conversion from sodium sulfite to vitamin C failed to obtain the genomic DNA from the samples. 5)When the filter with the blood cells was stored for four weeks at room temperature, genomic DNA was purified. We purified the genomic DNAs using this method for 10 yen/sample and from 16 samples at 1 hour. These results suggest that this method is a cost-effective and rapid genomic DNA purification method in animals and birds.
2010-03-16