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Use the NEB Tm Calculator to estimate an appropriate annealing temperature when using NEB PCR products.
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.
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™.
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.
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.
