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===Calculation=== Probe:target ratio 200:1 (according to Alice, 100:1 works too) *Billy uses 250ng NIH_3T3_gDNA Template(100ng/uL) Use 200ng total, so = 2uL *One genome mass = 3pg *First calculate the # of genome your target represents *For 200ng, it represents 200ng/3pg=66666.67 genomes *You want to have 200x the probes as the target 200ng/3pg*200 *You want every single probe to target ratio to be 200:1 200ng/3pg*200*55,000 Ms_Meth 22ng/uL 55,000Probes 100nt Since 1 base = 330Da (for single stranded DNA, 660Da for ds), the concentration of probe is 22 ng/ul /(100x330) = 0.000666667 = 666.67nM # of molecules per mole = 6X10^23 # of probes needed/ concentration of probes = Volume of probes needed the volume the probe needed is 200ng/3pg x 200 x 55,000 / [(6x10^23)x (666.67*10^-9M)]*10^6 =1.83uL
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