An inexpensive, simple protocol for DNA isolation from blood for high-throughput genotyping by polymerase chain reaction or restriction endonuclease digestion.
We describe simple, inexpensive, and reliable methods for isolating DNA from avian blood, semen, or feather pulp. The procedures are readily applicable to high-throughput 96-well plate isolation for genotype analysis of chicken DNA based on restriction endonuclease digestion or PCR. Isolation cost is primarily the cost of a deep-well assay block and a few pipet tips; current price is less than 0.10 dollar per sample, providing a significant cost advantage over commercial kits. The procedure employs inexpensive, nonhazardous reagents and yields intact, double-stranded DNA from as little as 2 to 10 microL of avian blood, suitable for RFLP analysis or hundreds of PCR amplifications. We compared our method to published procedures for alkaline extraction from feather pulp and found our method to be more reliable with the advantage of isolating intact DNA sequences that can be easily quantified. With minor modifications, the method can isolate DNA for PCR genotyping from mammalian whole blood.
Bailes
S M
SM
Missouri State Highway Patrol Crime Laboratory, Jefferson City 65101, USA.
Devers
J J
JJ
Kirby
J D
JD
Rhoads
D D
DD
D000818:Animals; D002645:Chickens / Q000235:genetics*; D004247:DNA / Q000097:blood* / Q000302:isolation & purification*; D005838:Genotype; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods / Q000662:veterinary*; D015183:Restriction Mapping / Q000191:economics / Q000379:methods / Q000662:veterinary*
D004247:DNA