What types of templates can the Sequencing Core process?

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

 


For EACH kb of fragment size, 3 ng/ul10 ul per reaction1 pMol/ul10 ul per reactionImportant! Remove old PCR primers!!
Large DNA (plasmids over ~20 kb, cosmids, lambda clones)500 ng/ul10 ul per reaction30 pMol/ul (30 uM)20 ul per reactionCaution! Experimental! Talk to the Core Director.
DNA fragments  (e.g. gel isolated fragments)For EACH kb of fragment size, 3 ng/ul10 ul per reaction1 pMol/ul10 ul per reaction 
Single Stranded (e.g. M13)25 ng/ul10 ul per reaction1 pMol/ul10 ul per reaction 
Plasmids (mini-preps, maxi-preps, 4-15kb)50 ng/ul10 ul per reaction1 pMol/ul10 ul per reaction 
Large DNA  (P1 clones, BACs, bacterial genomic)1 ug/ul  Yes, really!10 ul per reaction30 pMol/ul (30 uM)20 ul per reactionCaution! Experimental! Talk to the Core Director.
siRNA Constructs200 ng/ul10 ul per reaction3 pMol/ul20 ul per reaction 
Rolling-Circle preps  (e.g. 'TempliPHI' or equivalent)As per manufacturer's recommendation  (typ 10X dilution of product)10 ul per reaction1 pMol/ul10 ul per reaction 

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