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Use the NEB Tm Calculator to estimate an appropriate annealing temperature when using NEB PCR products.
OligoAnalyzer™ Tool. Understand the expected properties of your oligos before you order them. Calculator for GC content, melting temperature (T m), molecular weight, extinction coefficient, µg/OD, nmol/OD, and more. Identify secondary structure potential. Minimize dimerization.
The calculator calculates recommended T m (melting temperature) of primers and PCR annealing temperature based on the primer pair sequence, primer concentration, and DNA polymerase used in PCR. The calculator also calculates the primer length, percentage of GC content, molecular weight, and extinction coefficient.
8 lis 2021 · PCR primers are interdependent with your sample type, target, reagents, and thermodynamic conditions. Establishing your PCR objective can guide your primer design and enzyme choices. It’s a good idea to review parameters and understand why primer design is such a critical, dependent variable in PCR.
15 cze 2012 · OligoAnalyzer calculates Tm, molecular weight, hairpins, self-dimers, heterodimers, BLAST search, and Tm mismatch for custom DNA or RNA oligos.
Whether you are creating an assembly that incorporates multiple fragments, conducting your entire PCR at a single temperature, or incorporating intentionally mismatched nucleotides, careful primer design can set you up for success before you even pick up your pipette.
24 sie 2015 · 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.