A primer is a short single-stranded nucleic acid utilized by all living organisms in the initiation of DNA synthesis. The enzymes responsible for DNA replication, DNA polymerases, are only capable of adding nucleotides to the 3’-end of an existing nucleic acid, requiring a primer be bound to the template before DNA polymerase can begin a complementary strand. Living organisms use solely RNA primers, while laboratory techniques in biochemistry and molecular biology that require in vitro DNA synthesis (such as DNA sequencing and polymerase chain reaction) usually use DNA primers, since they are more temperature stable.
Dear Mohammed
in case it could be useful, i uploaded some video with guidelines for primer design for cloning of pcr fragments into plasmids at PAGE1 of my blog ProteoCool (https://proteocool.blogspot.com)
ciao
Manuele
PCR Primer Design (Methods in Molecular Biology) 2nd ed. 2015 Edition
Recombinant DNA: A Short Course, second (1992) or third (2006) edition. The ones written by James Watson, Jan Witkowsky and co.
Hey,
Use this link. It is easy to read and so far give the best result for primer designing!
http://biotools.nubic.northwestern.edu/OligoCalc.html
I think designing manually gives you a greater understanding of the molecular biology than following the book pattern.
Dear researcher,
You can check books like PCR Primer Design by Yuryev, Anton (2007) or
CR Primer Design by Basu, Chhandak (2015) all can be read in Methods in Molecular Biology.
You may have to read ''Understanding PCR: A Practical Bench-Top Guide'' written by Sarah Maddocks and Rowena Jenkins (2016). The first edition is a user friendly book that has step-by-step basic protocols, which can be adapted to your needs.
Goodluck.
snapgene or neb builder webpage is the best way to design primers, making them by hand is bad practice
Check: PCR Protocols - A Guide to Methods and Application; edited by, Michael A. Innis, David H. Gelfand, John J.Sninsky, Thomas J. White. Published by Academic Press.
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