Question

2.9. Find the volumes of each component for a PCR master mix for 18 reactions.

                                                                                                                        

No of reactions to make up			18
Vol of each Reaction (µL)			24 µL +1 µL DNA
Total Volume Required			µL total
Component	Supplied as	Final Conc in PCR	µL required 1rxn     18 rxn’s
MgCl2	25 mM	2.5 mM
Polymerase buffer	5x	1x
Primer forward (Bird F1)	10 µm	1 µm
Primer Reverse (Bird R1)	10 µm	1 µm
Taq DNA polymerase	1Units/µL	1 U per rxn
dNTPs	25 mM	1 mm
Template DNA*added later	-	-	1 µL *	18 µL *
Water	-	-
		Total volume	25 µL

Answer 1

No of reactions to make up	18
Vol of each Reaction (µL)	24 µL+1 µL DNA
Total Volume Required	25 µL total
Component	Supplied as	Final Conc in PCR	µLrequired 1rxn     18 rxn’s
MgCl2	25 mM	2.5 mM	2.5      45
Polymerase buffer	5x	1x	5          90
Primer forward (Bird F1)	10 µm	1 µm	2.5       45
Primer Reverse (Bird R1)	10 µm	1 µm	2.5       45
Taq DNA polymerase	1Units/µL	1 U per rxn	1     1
dNTPs	25 mM	1 mm	1     1
Template DNA*added later	-	-	1         18
Water	-	-	10       180
Total volume			25 µL   450 µL