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NEB Tm Calculator (tmcalculator.neb.com)
TmCalculator

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version 1.16.6
HELP    ABOUT   

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

Instructions

 1. Select the product group of the polymerase or kit you plan to use.
 2. Select the polymerase or kit from the list of products.
 3. If needed, modify the recommended primer concentration.
 4. Enter primer sequences (with up to 3 ambiguous bases). Spaces allowed.

Note that an anealing temperature will only be displayed if both primer
sequences are entered.

Product Group
Q5Q5 Hot StartQ5U Hot StartQ5 Blood DirectOneTaqOneTaq Hot StartHot Start TaqTaq
DNA PolymeraseLongAmp TaqLongAmp Hot Start TaqHemo KlenTaqVentDeep VentMaster
MixPhusionPhusion Hot Start FlexEpiMark Hot Start
Polymerase/Kit
Q5 High-Fidelity DNA PolymeraseQ5 High-Fidelity 2X Master Mix

Primer Concentration (nM)

Reset concentration
Primer 1
Primer 2


Switch to batch mode
Clear
Use example input

Anneal at



--- °C

Why is this so high?
Primer 1
--- nt
---% GC
Tm: ---°C
Primer 2
--- nt
---% GC
Tm: ---°C
Show expanded primer 1 sequences


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Primer 1 expanded (--- nt, Tm: ---°C )

Seq Tm %GC


Show expanded primer 2 sequences


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Primer 2 expanded (--- nt, Tm: ---°C)

Seq Tm %GC


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