I need a primer for some genes without introns and also it should check the DNA contamination to ensure there is no DNA contamination.
For normal genes we can use intron junctions to check (or control) the contaminations.
Hi Aref
It is better to put a -RT control to see if there is some residual DNA contamination left in your RNA preparations.
bye
you can use exon junction of Primer-BLAST(http://www.ncbi.nlm.nih.gov/tools/primer-blast/index.cgi?LINK_LOC=BlastHome).
As Ajay mentioned above, only possibility is extra controls and addl dnase treatment before RT.
Parisa farrokh
neither there is intron nor exon junction possibility...
I use the Primer-blast tool on NCBI to design intron-spanning primers by selecting the option under "Exon/Intron Selection" (See attachment). This ensures that the primers do not bind to DNA.
use a reference gene for your sample. So perform for your cDNA PCR with primers for other gene which has a intron and you know exactly how long is the product where the tamplate is genomic DNA and cDNA.
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