Band-cutting no more: A method for the isolation and purification of target PCR bands from multiplex PCR products using new technology.
A procedure for the isolation of target PCR bands from multiplex PCR products using the E-Gel iBase Power System, an E-Gel Safe Imager Real-Time Transilluminator, and E-Gel CloneWell 0.8% SYBR Safe agarose cassettes is presented. The collected isolates are suitable for direct use in sequencing reactions without need of further purification. Reductions in time spent per sample and cost per sequence produced compared to traditional "band-cutting" methods are demonstrated. An added benefit is that the new procedure does not require exposure to ethidium bromide or ultraviolet radiation.
Gibson
Joel F
JF
Agriculture and Agri-Food Canada, Canadian National Collection of Insects, Arachnids and Nematodes, 960 Carling Avenue, Ottawa, ON, Canada K1A 0C6. joel.gibson@agr.gc.ca
Kelso
Scott
S
Skevington
Jeffrey H
JH
D009696:Nucleic Acids / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D017421:Sequence Analysis / Q000379:methods*
D009696:Nucleic Acids