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Use the NEB Tm Calculator to estimate an appropriate annealing temperature when using NEB PCR products.
Use this tool for your scientific calculations and conversions for DNA and RNA. Options include conversion of mass to moles, ligation amounts, conversion of OD to concentration, dilution and molarity. Additional features include sgRNA Template Oligo Design and qPCR library quantification.
Use this tool to estimate an appropriate incubation temperature when using NEB thermostable ligases and reaction buffers.
The NEB Tm Calculator is recommended to calculate an appropriate annealing temperature. When primers with annealing temperatures above 65°C are used, a 2-step PCR protocol is possible (see #10). Extension: The recommended extension temperature is 68°C. Extension times are generally 1 minute per kb.
28 lip 2022 · Use Tm Calculator to calculate optimum annealing temperature for PCR primers when using NEB polymerases and buffers. Use NEBioCalculator to convert DNA or RNA mass to moles, calculate...
The NEB Tm calculator should be used to determine the annealing temperature when using Phusion. Typically, primers greater than 20 nucleotides in length anneal for 10–30 seconds at 3°C above the T m of the lower T m primer.
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