In vitro selection of RNA aptamers to a protein target by filter immobilization.
This unit describes the selection of aptamers from a pool of single-stranded RNA by binding to a protein target. Aptamers generated from this selection experiment can potentially act as protein function inhibitors, and may find applications as therapeutic or diagnostic reagents. A pool of dsDNA is used to generate an ssRNA pool, which is mixed with the protein target. Bound complexes are separated from unbound reagents by filtration, and the RNA:protein complexes are amplified by a combination of reverse transcription, PCR, and in vitro transcription.
Hall
Bradley
B
Department of Chemistry and Biochemistry, University of Texas, Austin, Texas, USA.
Arshad
Seyed
S
Seo
Kyunghyun
K
Bowman
Catherine
C
Corley
Meredith
M
Jhaveri
Sulay D
SD
Ellington
Andrew D
AD
D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism*; D005374:Filtration / Q000379:methods*; D007202:Indicators and Reagents; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000235:genetics / Q000378:metabolism*
D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D007202:Indicators and Reagents; D011506:Proteins