Gibco Taq or Platnium DNA Polymerase Data Sheet FGB

Gibco BRL: 10966-018 10/2/98

Date:________ Kit used:________ Oligo’s used:_________

Samples:_____________________________________________

PCR Reaction:

Recommended ul per reaction: ul used: Master Mix: PCR Cycles: Time\Temp:

10X PCR Buffer __5__ _____ _____ One Denaturation:

Annealing:

50. mM MgCl2 __1.5_ _____ _____ Extension:

10 mM dNTP’s __1___ _____ _____ ____ Denaturation:

Annealing:

10uM 3’ oligo __1___ _____ _____ Extension:

10uM 5’ oligo __1___ _____ _____

18S Standard 0.3:0.7 _____ _____

DNA __1-10 _____ _____

Sterile H2O up to 50ul _____ _____

Platinum Taq __0.5__ _____ _____

(or Gibco Taq)

___________________________________

F.V.= 50. use _____ ul of master mix per sample.

Final concentrations: 1x buffer; 0.2mM each dNTP; 1.5mM MgCl2; 0.2uM each primer; 10-100ng DNA; 2.5units Taq or as required.

1. A final volume of 50 ul per sample is suggested.

2. On ice, prepare a master mix for one more sample than you are planning to perform.

3. Include all the reagents that are not variables (i.e. RT mix or MgCl2 usually).

4. Bring up to 50 ul then mix well and spin down.

5. On ice, overlay with sterile mineral oil if needed then run PCR.

6. Run 8-10 ul of your PCR products in a 1-1.5% gel with EtBr in 0.5X TBE at 100v.