Login
Password

Forgot your password?

Real Time PCR Primer Design

By Edited Nov 26, 2016 0 0

It is important that real time PCR primers are designed properly. Poorly designed primers will contain complementary sequences often binding on themselves and resulting in primer dimer and poor annealing to DNA template. Hence proper design of primers will save time, effort and money.


Follow the simple steps to design a primer


1. Go to Pubmed.com and enter the details of the gene of interest and choose "Nucleotide" in the drop down menu.


2. Look for the complete CDS or the mRNA sequence.


3. Copy the open reading frame sequence into word and highlight the regions between the introns and exons. Remember that when introns are removed the exons can be brought together. This is the region that is made into protein.


Alternatively, intron exon boundaries can be identified using a programme that compares the given genomic DNA sequence with the cDNA. Usually such a programme colours the 2 sections and then the introns can be removed.


4. Finally the PCR primes can be designed against 2 exon boundaries.

Advertisement

Comments

Add a new comment - No HTML
You must be logged in and verified to post a comment. Please log in or sign up to comment.

Explore InfoBarrel

Auto Business & Money Entertainment Environment Health History Home & Garden InfoBarrel University Lifestyle Sports Technology Travel & Places
© Copyright 2008 - 2016 by Hinzie Media Inc. Terms of Service Privacy Policy XML Sitemap

Follow IB Technology