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

  2. Use this tool when designing PCR reaction protocols to help determine the optimal annealing temperature for your amplicon. Simply input your DNA polymerase, primer concentration and your primer sequence and the Tm Calculator will guide you to successful reaction conditions.

  3. 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.

  4. www.neb.com › en › neb-primer-design-toolsNEB ® Primer Design Tools

    NEBaseChanger can be used to design primers specific to the mutagenesis experiment you are performing using the Q5 Site-Directed Mutagenesis Kit. This tool will also calculate a recommended custom annealing temperature based on the sequence of the primers by taking into account any mismatches.

  5. Tm Calculator. Use this tool when designing PCR reaction protocols to help determine the optimal annealing temperature for your amplicon. Simply input your DNA polymerase, primer concentration and your primer sequence and the Tm Calculator will guide you to successful reaction conditions.

  6. NEB Tm Calculator is used to estimate an appropriate annealing temperature in NEB PCR products. The tool fields are: Product Group: select a value from Q5, Q5 Hot Start, Q5U Hot Start, OneTaq, OneTaq Hot Start, Hot Start Taq, Taq DNA Polymerase, LongAmp Taq, LongAmp Hot Start Taq, Hemo KlenTaq, Vent, Deep Vent, Master Mix, Phusion, Phusion Hot ...

  7. 27 lip 2023 · Please use NEB’s Tm Calculator to determine the appropriate annealing temperature for your primer pair and NEB polymerase/buffer of interest. Unlike other calculators, the NEB Tm Calculator takes into consideration buffer components that affect melting temperatures and empirical observations when calculating the optimal annealing temperature.

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