Different substances such as dimethyl sulfoxide, tetramethylene sulfoxide, 2-pyrollidone, and the naturally occurring compatible solute betaine enhance PCR amplification of GC-rich DNA templates with high melting temperatures. In particular, cyclic compatible solutes outperform traditional PCR enhancers. We therefore investigated the effects that cyclic naturally occurring ectoine-type compatible solutes and their synthetic derivatives have on melting temperature of double-stranded DNA (dsDNA) and on PCR amplification of different templates. L-ectoine, betaine, and derivatives of L-ectoine decreased, whereas beta-hydroxyectoine increased, the melting temperature of dsDNA. The ability to decrease the melting temperature was greatest for homoectoine, a new synthetic derivative of l-ectoine. Furthermore, compatible solutes, especially homoectoine, enhanced PCR amplification of GC-rich DNA (72.6% GC content; effective range: 0.1-0.5M).
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1016/j.bbrc.2004.07.200 | DOI Listing |
Foods
December 2024
School of Public Health, Hebei Medical University, Shijiazhuang 050017, China.
Foodborne viruses are significant contributors to global food safety incidents, posing a serious burden on human health and food safety. In this study, a multiplex reverse transcription-droplet digital PCR (RT-ddPCR) assay based on the MS2 phage as a process control virus (PCV) was developed to achieve the simultaneous detection of hepatitis A virus (HAV) and hepatitis E virus (HEV) in bivalve shellfish. By optimizing the reaction system and procedures, the best reaction conditions were selected, and the specificity, sensitivity, and reproducibility of the method were assessed.
View Article and Find Full Text PDFAnimals (Basel)
December 2024
State Key Laboratory of Swine and Poultry Breeding Industry, National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
Flamingos () are among the oldest birds worldwide and are loved by people for their bright red feathers. In addition, flamingos are sexually monomorphic birds, and distinguishing between males and females is difficult. The polymerase chain reaction (PCR) is widely used for sex identification.
View Article and Find Full Text PDFPlanta
January 2025
Advanced Laboratory for Plant Genetic Engineering, Advanced Technology Development Centre, Indian Institute of Technology, Kharagpur, India.
This study seeks to improve the biomass extractability of Sorghum bicolor by targeting a critical enzyme, 4CL, through metabolic engineering of the lignin biosynthetic pathway at the post-transcriptional level. Sorghum bicolor L., a significant forage crop, offers a potential source of carbohydrate components for biofuel production.
View Article and Find Full Text PDFSci Rep
January 2025
Institute for Biosecurity and Microbial Forensics (IBMF), Oklahoma State University, Stillwater, OK, USA.
Exclusion is a keystone of integrated pest management to prevent the introduction of pathogens. U.S.
View Article and Find Full Text PDFClin Microbiol Infect
January 2025
Chlamydia Group, Institute of Veterinary Pathology; University of Zürich, Switzerland.
Objectives: Chlamydia trachomatis is the most commonly diagnosed bacterial sexually transmitted infection (STI) worldwide. Diagnosis relies on nucleic acid amplification techniques, such as PCR, which does not distinguish between viable pathogens and residual bacterial DNA, leading to potential overdiagnosis and overtreatment. PCR with confirmation of pathogen viability has not been widely explored in the STI field.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!