I designed a primer for vancomycin resistant gene Van A ,when I made PCR there was no result and all sample showed A primer dimer,so I guess that my primer design is incorrect ,how can i check this??
You could check your primer pair at the primer3 website:
http://primer3.ut.ee/
To find out if your primers are binding at the right position on your template, you could apply BLASTn. It will reveal the expected binding positions and indicate the orientation of the binding as well. This is especially useful to check the orientation of the reverse primer.
One way of minimising primer dimer is to use a hot start enzyme. Once p-d starts it is very efficient and very often you will get no product because all of the primer is removed. With hot start the small regions of primer overlap never get a chance to anneal because the pcr temperature never drops below annealing temperature when there is active enzyme present.
Perform Primer BLAST at https://www.ncbi.nlm.nih.gov/tools/primer-blast/
If you get the desired amplicon and gene as a result of primer blast, primers designed are correct. Then you further need to optimize PCR from all aspect right from annealing temperature to conc of primers, template, Mg etc.
You can use IDT oligo analyser for primer designing, specifying the primer base no., Tm, GC content etc. Choose a primer set with minimum secondary structure formation. Thereafter can use Primer BLAST for the primers obtained using IDT oligo analyser. If the blast result shows the the genetic sequence of your desired gene, the primers designed are perfect.
I use PRIMER blast, Gene Runner and OLIGO to design many primers, and the results have been good.They indicate the possibility of the formation of structures such as dimer primer, hairpin, etc.I suggest you check your primer pairs with these tools before synthesizing the primers.