A simple method for sequencing small DNAs by introducing precise overlapping ends into restriction digestion fragments.
A method has been devised whereby certain complete restriction digestions can have short overlaps of unique sequence incorporated into the fragments during cloning. Thus one can identify when the DNA fragments are contiguous. Here, this technique is used to produce a contig for a 2.5 kb fragment of genomic DNA. This is a simple approach to ordering digestion fragments without necessarily performing restriction mapping. It is envisaged that this technique will be useful for sequencing cDNAs and small genomic fragments.
Division of Biochemistry and Molecular Biology, Davidson Building, IBLS, University of Glasgow, Glasgow G12 8QQ, UK. grena@bad.dundee.ac.uk
G
G
Rena
M D
MD
Houslay
D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D004262:DNA Restriction Enzymes; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D016678:Genome; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods*
D017931:DNA Primers; D018076:DNA, Complementary; D004247:DNA; D004262:DNA Restriction Enzymes