Long PCR
Definition of Long PCR:
Long PCR is a PCR is which extended or longer than standard PCR, meaning over 5 kilobases (frequently over 10 kb). Long PCR is usually only useful if it is accurate. Thus, special mixtures of proficient polymerases along with accurate polymerases such as Pfu are often mixed together.
Applications of Long PCR
Long PCR is often used to clone larger genes or large segments of DNA which standard PCR cannot.
External Links to Long PCR Articles
Long PCR Reagents and Guidelines
See LA PCR (Long and Accurate PCR)
Long PCR Protocols
Optimization and Tips for Long PCR
Long PCR Amplification Procedure
Long PCR Recently Published Research
A Rapid Long PCR-Direct Sequencing Analysis for ABO Genotyping.
A Rapid Long PCR-Direct Sequencing Analysis for ABO Genotyping.
Ann Clin Lab Sci. 2011;41(4):340-5
Authors: Huh JY, Park G, Jang SJ, Moon DS, Park YJ
Abstract
ABO is the most clinically important blood group system in transfusion and transplantation medicine. The popular ABO genotyping methods, such as the sequencing of exons 6 and 7 and sequence-specific primer (SSP)-PCR, often lead to ambiguous typing results. Long PCR-sequencing method was designed to analyze two regulatory regions (promoter and CBF/NF-Y enhancer regions) and all genomic sequences (except for intron 1) of the ABO gene. Using rapid DNA polymerase with high-fidelity, we amplified 6.3 kb and 7.3 kb for sequencing of enhancer-exon 1 and exons 2-7, respectively. ABO genotyping was performed using this technique in the peripheral blood of three unrelated families. The time requirements of the PCR amplification and purification processes were about 2.0 hours and 15 minutes, respectively. Five different ABO alleles (ABO*A102, ABO*A105, ABO*O01, ABO*O02, and ABO*B101) with allele-specific CBF/NF-Y minisatellite repeats from three families were analyzed. All genotyping results agreed with serologic findings and results expected by Mendelian inheritance. Compared to conventional PCR-direct sequencing for ABO genotyping, this method proves simple and fast for the analysis of ABO genotypes. Therefore, it might be valuable in clinical transfusion or forensic applications.
PMID: 22166503 [PubMed - in process]