TY - JOUR
T1 - Identifying transcribed sequences in arrayed bacteriophage or cosmid libraries.
AU - Hochgeschwender, U.
AU - Brennan, M. B.
PY - 2001/5
Y1 - 2001/5
N2 - This unit describes methods for identifying bacteriophage or cosmid clones that contain sequences present in the mRNA of a cell line or tissue. In the , a radiolabeled cDNA probe is synthesized by reverse transcription of poly(A)+ RNA isolated from the cell line or tissue of interest. The probe is incubated with DNA-cellulose to remove highly repetitive sequences before it is used for hybridization analysis of filters from a phage or cosmid genomic library. After identification of positive clones from the library screen, the same probe can be used to screen Southern blots of restriction enzyme-digested DNA from the positive clones. Support protocols describe preparation and testing of DNA-cellulose.
AB - This unit describes methods for identifying bacteriophage or cosmid clones that contain sequences present in the mRNA of a cell line or tissue. In the , a radiolabeled cDNA probe is synthesized by reverse transcription of poly(A)+ RNA isolated from the cell line or tissue of interest. The probe is incubated with DNA-cellulose to remove highly repetitive sequences before it is used for hybridization analysis of filters from a phage or cosmid genomic library. After identification of positive clones from the library screen, the same probe can be used to screen Southern blots of restriction enzyme-digested DNA from the positive clones. Support protocols describe preparation and testing of DNA-cellulose.
UR - http://www.scopus.com/inward/record.url?scp=44049102903&partnerID=8YFLogxK
U2 - 10.1002/0471142905.hg0602s00
DO - 10.1002/0471142905.hg0602s00
M3 - Article
C2 - 18428298
AN - SCOPUS:44049102903
SN - 1934-8258
VL - Chapter 6
SP - Unit 6.2
JO - Current protocols in human genetics / editorial board, Jonathan L. Haines ... [et al.]
JF - Current protocols in human genetics / editorial board, Jonathan L. Haines ... [et al.]
ER -