PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates.
AUTOR(ES)
Barnes, W M
RESUMO
A target length limitation to PCR amplification of DNA has been identified and addressed. Concomitantly, the base-pair fidelity, the ability to use PCR products as primers, and the maximum yield of target fragment were increased. These improvements were achieved by the combination of a high level of an exonuclease-free, N-terminal deletion mutant of Taq DNA polymerase, Klentaq1, with a very low level of a thermostable DNA polymerase exhibiting a 3'-exonuclease activity (Pfu, Vent, or Deep Vent). At least 35 kb can be amplified to high yields from 1 ng of lambda DNA template.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=43341Documentos Relacionados
- Induction of human papillomavirus type 18 late gene expression and genomic amplification in organotypic cultures from transfected DNA templates.
- Fingerprinting of diverse species with DNA probes generated from immobilized single-stranded DNA templates.
- Generation of labeled RNA probes from enzymatically amplified DNA templates.
- RNA polymerase of Myxococcus xanthus: purification and selective transcription in vitro with bacteriophage templates.
- A mathematical model and a computerized simulation of PCR using complex templates.