I want the best program for determine annealing temperature and band size for the primer?

There are several software programs available for determining the annealing temperature and band size for PCR primers. Here are a few of the best options:

Primer3: This is a free, open-source program that is widely used for designing PCR primers. It allows you to input your target DNA sequence and design primers based on a variety of parameters, including annealing temperature, GC content, and primer length. Primer3 also includes several useful features, such as the ability to design primers for multiplex PCR reactions and the option to include restrictions enzyme sites in your primers.

OligoCalc: This is another free software program that is widely used for primer design. It allows you to input your target DNA sequence and design primers based on several parameters, including annealing temperature, GC content, and melting temperature. OligoCalc also includes several advanced features, such as the ability to design primers for real-time PCR and the option to check for potential primer-dimer formation.

Geneious Prime: This is a commercial software program that includes a wide range of tools for molecular biology research, including primer design. It allows you to input your target DNA sequence and design primers based on a variety of parameters, including annealing temperature, GC content, and primer length. Geneious Prime also includes several advanced features, such as the ability to design primers for sequencing and the option to optimize primer design for specific PCR conditions.

Ultimately, the best program for determining annealing temperature and band size for PCR primers will depend on your specific needs and preferences. It's a good idea to try out several different programs and see which one works best for your particular application.

Here some videos about primer design with might be helpful:

How to design primers for PCR https://youtu.be/V8oG1BMRT2k

Forward and reverse primers https://youtu.be/BKpCdxohFZM

Forward and reverse primers https://youtu.be/Zbx7acBmAB4

How to choose Primers for PCR https://youtu.be/RaKSF4rZaZ8

Forward and reverse primers explained https://youtu.be/L8oAnxWoN-w

Polymerase chain reaction and primers design https://youtu.be/TT-bE1xkPSs

How to choose primers https://youtu.be/Y63TDfKcrDU

Primers, Okazaki fragments, mRNA, nicks, DNA Supercoiling https://youtu.be/2ssQ5-gPW4M

How to design primers for PCR https://youtu.be/UUbL7u7SIxY

How to design primers https://youtu.be/FWRxXcoM0hk

Forward and reverse, sense and antisense primers https://youtu.be/0vdbf4F90_o

Sabine Strehl

Here are some popular online tools:

https://www.ncbi.nlm.nih.gov/tools/primer-blast/

https://primer3.ut.ee/

https://www.bioinformatics.nl/cgi-bin/primer3plus/primer3plus.cgi

Pakorn Pengin

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

https://tmcalculator.neb.com/

Net Primer: Free! Primer Analysis Software

https://www.premierbiosoft.com/netprimer/

Bandla Ramesh

There are several programs available for determining annealing temperature and band size for primers. Some popular programs include Primer3 (http://primer3.ut.ee/), OLIGO (http://www.oligoweb.com/), and Primer-BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi?PROGRAM=blastn&PAGE_TYPE=BlastSearch&LINK_LOC=primer-blast).

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