Manganese citrate improves base-calling accuracy in DNA sequencing reactions using rhodamine-based fluorescent dye-terminators.
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.
Korch
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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. christopher.korch@uchsc.edu
Drabkin
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H
D001483:Base Sequence; D019343:Citric Acid / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D008969:Molecular Sequence Data; D009942:Organometallic Compounds / Q000737:chemistry*; D012235:Rhodamines / Q000737:chemistry*; D017422:Sequence Analysis, DNA / Q000592:standards*