A simple method for sequencing small DNAs by introducing precise overlapping ends into restriction digestion fragments
Graham Rena, Miles D. Houslay
Abstract
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Graham Rena, Miles D. Houslay
Abstract
Open-access reader
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.
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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.
Key concepts: Biology, Contig, Restriction digest, Restriction fragment, Restriction map, Restriction site, Restriction enzyme, Terminal restriction fragment length polymorphism