Field inversion gel electrophoresis applied to the rapid, multi-enzyme restriction mapping of phage lambda clones

We have developed digestion conditions, FIGE gel electrophoresis conditions, and a size standard for rapid multienzyme restriction mapping of inserted DNA in phage lambda vectors. lambda DNA (carrying cloned inserts) can be mapped by partially digesting it, hybridizing to a labeled oligonucleotide c...

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Bibliographic Details
Published in:Nucleic acids research 1990-03, Vol.18 (5), p.1312-1312
Main Authors: DANIELS, D. L, OLSON, C. H, BRUMLEY, R, BLATTNER, F. R
Format: Article
Language:English
Subjects:
Bacteriophage lambda - genetics
Biological and medical sciences
Electromagnetic Fields
Electrophoresis - methods
Electrophoresis, Agar Gel - methods
Fundamental and applied biological sciences. Psychology
Miscellaneous
Molecular and cellular biology
Molecular genetics
phage lambda
restriction endonuclease mapping
Restriction Mapping
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Description
Summary:We have developed digestion conditions, FIGE gel electrophoresis conditions, and a size standard for rapid multienzyme restriction mapping of inserted DNA in phage lambda vectors. lambda DNA (carrying cloned inserts) can be mapped by partially digesting it, hybridizing to a labeled oligonucleotide complementary to the single-stranded cos overhang at the end of lambda , separating the products by gel electrophoresis, and visualizing the result by autoradiography. This procedure is conceptually straightforward, but its use has been severely handicapped because of the difficulty of controlling the extent of digestion and the fact that the lambda ends are at some distance from the cloned insert.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/18.5.1312