2018年2月14日 · Probe and primer are two types of single-stranded, oligonucleotides used in various PCR techniques to hybridize with complementary DNA. Both probe and primer are specific to a particular DNA fragment.

2013年10月21日 · Many factors can influence PCR experiments, including primer and probe characteristics such as location, length, interaction and self-folding, melting temperature, annealing temperature, and GC content. Review these general recommendations for designing primers and probes and for choosing target locations for PCR amplification.

Learn about PCR primer and probe chemistries for real-time PCR. Discover DNA primers with fluorescent DNA-binding dyes, hydrolysis probes and molecular beacons.

Real-time PCR primer assays consist of unlabeled PCR primer pairs for use with dye-based chemistry such as SYBR Green or EvaGreen. Probe assays for real-time PCR and Droplet Digital PCR include PCR primers and a dual-labeled fluorescent probe with your choice of fluorophore.

2022年9月5日 · PCR primers should have a length between 18 and 24 nucleotides, and probes between 15 to 30 nucleotides. The optimal melting temperature (T m) of a primer is 54°C or higher. The annealing temperature (T a) of a primer is often above its Tm (of 2-5°C). The GC content of a primer should be between 40% and 60%.

Enter the target sequence in FASTA format or an accession number of an NCBI nucleotide sequence in the PCR Template section of the form. If the NCBI mRNA reference sequence accession number is used, the tool will automatically design primers that …

2015年8月24日 · Simplify planning of your qPCR experiments using IDT free, online tools for oligonucleotide analysis and PCR primer design. This article provides an overview of our predesigned qPCR sequences and the basics of designing customized PCR primers and hydrolysis probes with the PrimerQuest™ Tool.

Review options for custom-synthesized oligos, primers, probes, and genes for PCR, cloning, hybridization, real-time PCR, protein expression. Also, access our easy oligo ordering portal or primer and probe design tools.

Optimal primer sequences and appropriate primer concentrations are essential for maximal specificity and efficiency in PCR. The table, Primer design and usage guidelines, provides an overview of primer design and use for standard and multiplex PCR, as well as one-step RT-PCR.

2015年1月1日 · Design of primers and probes is one of the most crucial factors affecting the success and quality of quantitative real-time PCR (qPCR) analyses, since an accurate and reliable quantification depends on using efficient primers and probes.