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Method for construction of normalized cDNA libraries

Patent ·
OSTI ID:871950
 [1];  [2]
  1. New York, NY
  2. Englewood, NJ
+ Show Author Affiliations

This invention provides a method to normalize a directional cDNA library constructed in a vector that allows propagation in single-stranded circle form comprising: (a) propagating the directional cDNA library in single-stranded circles; (b) generating fragments complementary to the 3' noncoding sequence of the single-stranded circles in the library to produce partial duplexes; (c) purifying the partial duplexes; (d) melting and reassociating the purified partial duplexes to appropriate Cot; and (e) purifying the unassociated single-stranded circles, thereby generating a normalized cDNA library. This invention also provides normalized cDNA libraries generated by the above-described method and uses of the generated libraries.

Research Organization:
Columbia University
DOE Contract Number:
FG02-91ER61233
Assignee:
Trustees of Columbia University in City of New York (New York, NY)
Patent Number(s):
US 5830662
OSTI ID:
871950
Country of Publication:
United States
Language:
English

References (3)

Construction of a uniform-abundance (normalized) cDNA library. journal March 1991
Construction of a normalized cDNA library by introduction of a semi-solid mRNA-cDNA hybridization system journal January 1994
An ‘equalized cDNA library’ by the reassociation of short double-stranded cDNAs journal January 1990

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Related Subjects

/435/999/
above-described
allows
allows propagation
appropriate
appropriate cot
cdna
cdna libraries
cdna library
circle
circle form
circles
coding sequence
complementary
comprising
constructed
construction
cot
directional
directional cdna
duplexes
form
form comprising
fragments
fragments complementary
generated
generating
generating fragments
libraries
library
library constructed
melting
method
noncoding
noncoding sequence
normalize
normalized
normalized cdna
partial
partial duplexes
produce
produce partial
propagating
propagation
provides
purified
purified partial
purifying
reassociating
sequence
single-stranded
single-stranded circle
single-stranded circles
unassociated
unassociated single-stranded
vector