Template Type: Template Conc: Template Amt: Primer Conc: Primer Amt: Notes:
Bacterial Genomic DNA We have had some limited success with sequencing from complete bacterial genomic DNA. Ask the Director about this before submitting. 10 ul per reaction   per reaction Caution! Experimental! Talk to the Core Director.
PCR products (rule of thumb)

 

For EACH kb of fragment size, 3 ng/ul 10 ul per reaction 1 pMol/ul 10 ul per reaction Important! Remove old PCR primers!!
Large DNA (plasmids over ~20 kb, cosmids, lambda clones) 500 ng/ul 10 ul per reaction 30 pMol/ul (30 uM) 20 ul per reaction Caution! Experimental! Talk to the Core Director.
DNA fragments  (e.g. gel isolated fragments) For EACH kb of fragment size, 3 ng/ul 10 ul per reaction 1 pMol/ul 10 ul per reaction  
Single Stranded (e.g. M13) 25 ng/ul 10 ul per reaction 1 pMol/ul 10 ul per reaction  
Plasmids (mini-preps, maxi-preps, 4-15kb) 50 ng/ul 10 ul per reaction 1 pMol/ul 10 ul per reaction  
Large DNA  (P1 clones, BACs, bacterial genomic) 1 ug/ul  Yes, really! 10 ul per reaction 30 pMol/ul (30 uM) 20 ul per reaction Caution! Experimental! Talk to the Core Director.
siRNA Constructs 200 ng/ul 10 ul per reaction 3 pMol/ul 20 ul per reaction  
Rolling-Circle preps  (e.g. 'TempliPHI' or equivalent) As per manufacturer's recommendation  (typ 10X dilution of product) 10 ul per reaction 1 pMol/ul 10 ul per reaction  

Please re-check this table periodically to see if we have adjusted our recommended concentrations.