Fluorescent and radioactive solid phase dideoxy sequencing of pcr products in microtitre plates.
In this paper we describe a rapid method for the direct generation of DNA sequencing templates from phage or bacteria. Sequencing of these PCR products can be performed by radioactive and fluorescent methods. The non-radioactive method has been used to sequence a total of approximately 100 kb of human DNA fragments generated by digestion with HpaII and subsequent cloning. The method depends on direct small scale amplification using a biotinylated primer, and the binding of the product to streptavidin coated magnetic beads. All the procedures are carried out in a microtitre plate thus facilitating the handling of large numbers of clones and has potential for automation.
Jones
D S
DS
MRC Molecular Genetics Unit, Cambridge, England.
Schofield
J P
JP
Vaudin
M
M
D001419:Bacteria / Q000235:genetics; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D001710:Biotin; D004247:DNA; D005453:Fluorescence; D005821:Genetic Techniques; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D013462:Sulfur Radioisotopes
D013462:Sulfur Radioisotopes; D001710:Biotin; D004247:DNA