|
Answer» You will get 100 micro molar final conc of each primers if you DISSOLVE in the appropriate volume they have mentioned in volume-pmol/microlitre. this will be your stock. but how i usually do is for mutagenesis i use 125 ng of primers in total. so i would suggest you to dissolve as they mentioned but calculate LIKE below, for Primer 1 in ur sheet: 594 microlitres will give you 100micromolar final concentration this is one way.... and the other is ,they also mentioned it is 805 micrograms so now if you dissolve 805 mcirograms in 594 volume (just not to alter the universal primer stock conc_100miromolar) this will give you-1.36 micrograms/microlitre (805/594), so now your stock is 1.36 microgram/microlitre or 100micromolar. to get 100ng/microlitre (WORKING) - 0.1/1.36x100 (total volume) = 7.35 so to prepare 100 ng/microlitre - 7.35 (stock primer)+ 92.65mcirolitres (H2O). now you can use 1.25 microlitres of each primers per pcr to get 125ng of each primer for a reaction. good luck....
|