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. 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.
Pollard
J
J
Harvard University, Cambridge, Massachusetts, USA.
Bell
S D
SD
Ellington
A D
AD
D000097:Acetonitriles; D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000138:chemical synthesis* / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009943:Organophosphorus Compounds / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D052156:SELEX Aptamer Technique / Q000379:methods*
D000097:Acetonitriles; D009943:Organophosphorus Compounds; C434331:phosphoramidite; D004247:DNA; C032159:acetonitrile