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Use the NEB Tm Calculator to estimate an appropriate annealing temperature when using NEB PCR products.
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. Use NCBI BLAST™.
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.
21 paź 2013 · The free, online IDT OligoAnalyzer Tool allows researchers to calculate T m for mismatched sequences. You can also use the program to determine probe locations that help enable better mismatch discrimination.
Browse NEB's beta tools, including REBSITES, Primer Monitor, Tm API, Glycan Analyzer Code and Genomes and Restriction Enzyme Frequencies, NEBNext Index Oligo Selector.
8 lis 2021 · Our Tm calculator accounts for specific buffer compositions that impact primer melting temps. The goal is to avoid weak thermodynamics that can cause primer dimers to form due to inefficient binding.
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.
