A rapid method for purifying PCR products for direct sequence analysis.
An HPLC approach for purification and sequencing of double-stranded DNA obtained directly from a PCR is described. This simple and reliable procedure has several advantages; the DNA fragment is rapidly eluted (less than 7 minutes), requires no organic cleanup, produces several hundred bases of sequence and is sensitive enough to obtain DNA sequence from a single 100-microliters PCR. This method is demonstrated by sequencing tumor necrosis factor alpha (TNF alpha) gene amplified from mouse tail DNA.
Department of Biological Sciences, Monsanto Company, St. Louis, MO 63198.
M H
MH
Kalnoski
M F
MF
McCoy-Haman
G F
GF
Hollis
D000818:Animals; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D051379:Mice; D008969:Molecular Sequence Data; D011089:Polydeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors
D011089:Polydeoxyribonucleotides; D004247:DNA