Long-range PCR remains a flexible, fast, efficient and cost-effective choice for sequencing candidate genomic regions in a small number of samples, especially when combined with next-generation sequencing (NGS) platforms. Several long-range DNA polymerases are advertised as being able to amplify up to 15 kb or longer genomic DNA. However, their real-world performance characteristics and their s...

This method for rapid, automated analysis of polymerase chain reaction (PCR) products makes use of PCR primers containing 5'-polypyrimidine sequences. Polypyrimidine-"headed" primers confer to the PCR product the ability to form triple helical complexes with a third polypyrimidine oligonucleotide. Third-strand oligonucleotides are modified to serve as either capture reagents or detection reagen...

BACKGROUND Haplotyping is an important technique in molecular diagnostics because haplotypes are often more predictive for individual phenotypes than are the underlying single-nucleotide polymorphisms (SNPs). Until recently, methods for haplotyping SNPs separated by kilobase distances were laborious and not applicable to high-throughput screening. In the case of thiopurine S-methyltransferase (...

In spite of controversial issues, pancreatectomy following neoadjuvant chemoradiation therapy (NeoCRT) has been applied in treating advanced pancreatic cancer. Cases of pathological complete remission (pCR) following NeoCRT is rare, and its long-term follow-up data are still lacking.From January 2000 to December 2012, medical records of the patients who underwent pancreatectomy for pancreatic d...

Neoadjuvant treatment with a sequential anthracycline-taxane-based chemotherapy in combination with trastuzumab (Herceptin) is currently a preferred therapy for patients with HER2-positive breast cancer. This approach is based on the higher pathologic complete response (pCR) of 40% seen with the addition of trastuzumab, compared with a 17% pCR with chemotherapy alone. The pCR can be increased t...

Accurate resolution of PCR products in the range of 15-40 kb may be obtained in agarose gels without pulsed field electrophoresis. A gel of 0.3% SeaKem Gold agarose cast on GelBond support film provides good resolution and sufficient get strength to reliably allow staining and photography. This paper describes a test system for Long PCR and demonstrates analysis of the PCR products on a gel run...

Formalin-fixed paraffin-embedded (FFPE) tissues are important resources for molecular medical research. However, long-chain RNA analysis is restricted in FFPE tissues due to high levels of degradation. To explore the possibility of long RNA quantification in FFPE tissues, we selected 14 target RNAs (8 mRNAs and 6 long noncoding RNAs) from literatures, and designed short (~60 bp) and long (~200 ...

BACKGROUND Familial hypercholesterolemia (FH) is caused by mutations in the low density lipoprotein (LDL) receptor gene. In this study we have compared multiplex ligation-dependent probe amplification (MLPA) and long-range PCR to detect large deletions/duplications in the LDL receptor gene. METHOD DNA from 431 unrelated FH patients without mutations in the LDL receptor gene detectable by DNA ...