Design, synthesis, and amplification of DNA pools for in vitro selection.
Preparation of a random-sequence DNA pool is presented. The degree of randomization and the length of the random sequence are discussed, as is synthesis of the pool using a DNA synthesizer or via commercial synthesis companies. Purification of a single-stranded pool and conversion to a double-stranded pool are presented as step-by-step protocols. Support protocols describe determination of the complexity and skewing of the pool, and optimization of amplification conditions.
Hall
Bradley
B
Department of Chemistry and Biochemistry, University of Texas, Austin, Texas, USA.
Micheletti
John M
JM
Satya
Pooja
P
Ogle
Krystal
K
Pollard
Jack
J
Ellington
Andrew D
AD
D004247:DNA / Q000302:isolation & purification / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000138:chemical synthesis* / Q000302:isolation & purification; D019020:Directed Molecular Evolution; D008967:Molecular Biology / Q000379:methods*; D011897:Random Allocation; D052156:SELEX Aptamer Technique / Q000379:methods*
D004277:DNA, Single-Stranded; D004247:DNA