What is primer melting point?

Primer melting temperature (Tm) by definition is the temperature at which one half of the DNA duplex will dissociate to become single stranded and indicates the duplex stability.

How do you calculate GC content in primers?

GC content is usually calculated as a percentage value and sometimes called G+C ratio or GC-ratio. GC-content percentage is calculated as Count(G + C)/Count(A + T + G + C) * 100%.

How high can primer Tm be?

As Mathis Wolter said the ideal TM of the primers must be between 50-60 C. Your TM is too high try to lower it down to at least 65-60 C, you can use temps of -5 C or + 5 C based on your Primers TM (i.e if you have a TM of 60C you can go either 55 C or 65 C).

What is the formula used in the Abi calculator?

The formula used in the ABI calculator is very simple. It goes as follows: Right ABI = highest right ankle systolic pressure / highest brachial systolic pressure Left ABI = highest left ankle systolic pressure / highest brachial systolic pressure Systolic blood pressure is the pressure on the walls of the blood vessels when the heart contracts.

What is the correct primer sequence to use for TM calculation?

Always use the actual primer sequence (i.e., 5′->3′ on minus strand of the template). Please enter the primer sequence only (No any other characters are allowed). The Tm calculation is controlled by Table of thermodynamic parameters and Salt correction formula (under advanced parameters).

What is the primer calculator used for?

The calculator also calculates the primer length, percentage of GC content, molecular weight, and extinction coefficient. The modified Allawi & SantaLucia’s thermodynamics method [1] is used for T m and annealing temperature calculation of reactions with Platinum SuperFi, Phusion and Phire DNA Polymerases.

What is the default BIP value for Primer3?

Schildkraut and Lifson 1965, DOI:10.1002/bip.360030207 (this is used until the version 1.0.1 of Primer3).The default value of Primer3 version 1.1.0 (for backward compatibility) 2. SantaLucia 1998, DOI:10.1073/pnas.95.4.1460 This is the recommended value.