柠檬酸锰可提高使用基于罗丹明的荧光染料终止剂的DNA测序反应中的碱基识别准确性。
Manganese citrate improves base-calling accuracy in DNA sequencing reactions using rhodamine-based fluorescent dye-terminators.
作者信息
Korch C, Drabkin H
机构信息
University of Colorado Cancer Center DNA Sequencing and Analysis Core and Division of Medical Oncology,School of Medicine, University of Colorado Health Sciences Center, Denver,CO 80262, USA.
出版信息
Nucleic Acids Res. 1999 Mar 1;27(5):1405-7. doi: 10.1093/nar/27.5.1405.
While dideoxy-terminators labeled with rhodamine-based fluorescent dyes provide the most versatile method of automated DNA sequencing, variation in peak heights reduces base-calling accuracy. We describe a simple approach that uses additions of a manganese salt and the metal buffer sodium citrate (MnCit) to overcome this limitation. This modification reduces peak height variability >2-fold and significantly increases the number of accurately read bases in DNA sequences.
虽然用罗丹明基荧光染料标记的双脱氧终止剂提供了最通用的自动化DNA测序方法，但峰高的变化会降低碱基识别的准确性。我们描述了一种简单的方法，即添加锰盐和金属缓冲剂柠檬酸钠（MnCit）来克服这一限制。这种改进使峰高变异性降低了2倍以上，并显著增加了DNA序列中准确读取的碱基数。
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