Simple preparation method of PCR fragments for automated DNA sequencing.
In an effort to find a simple and inexpensive purification method of polymerase chain reaction (PCR) reaction before cycle sequencing reaction, we compared a commercial system with a precipitation protocol performed in our laboratory. We found that, particularly with small PCR products, our method works with greater success than the method compared. Our precipitation method may be used on a larger PCR fragment before cycle sequencing reaction as well. Furthermore, it has the advantage of being simple as the well-known dilution method; in contrast to the dilution method, the precipitation method removes excess primers as well as possible primer dimers.
Høgdall
E
E
Department of Clinical Biochemistry, Statens Serum Institute, Copenhagen S, Denmark. eh@ssi.dk
Boye
K
K
Vuust
J
J
D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000235:genetics
D016159:Tumor Suppressor Protein p53; D004247:DNA