PCR Reagent Recipes

Avian PBS (Phosphate Buffered Solution)

1.08g Sodium phosphate dibasic (Na2HPO4) = di-sodium hydrogen orthophosphate

4.0g Sodium choloride (NaCl)

0.1g Potassium phosphate monobasic (KH2PO4)

0.1g Potassium cholride (KCl)

Top up to 500ml with ddH2O

Store in fridge

MgCl2 for PCR Reactions

1M MgCl2‚• 6H2O (100ml)

Dissolve 20.33g of MgCl2‚•H2O in 100ml of ddH2O

25mM MgCl2 PCR Solution

Add 25µl of 1M MgCl2 (from above) to 975µl of ddH2O

Freeze at -20˚C in 1.5ml microcentrifuge tubes


10X PCR Buffer

250ml of 1M KCl

25ml of 2M Tris (pH 8.8)

5ml of Triton X-100

8ml BSA (Bovine Serum Albumin)

Top up to 500ml with ddH2O


Aliquot into 1.5ml or 2.0ml microcentrifuge tubes and freeze at -20˚C

BSA (Bovine Serum Albumin)

• dissolve 0.8g of BSA in 8ml of water - mix and let dissolve over night in the fridge

Primer Dilutions

For desalted primers with the original concentration of 50nM , do the following to create a 20mM working primer stock:

• Amount of primer (in nm)/0.2 = x = amount of ddH20 added to tube to make a 200mM stock

• Dilute further to a microcentrifuge tube (20mM or 1/10 dilution) (20µl of 200mM primer +180µl of ddH20)

- this will be the working primer stock

* when diluting primers from the manufacturer, add the water and let sit for 30 minutes to an hour, vortex, and then make the dilutions (that way all of the primer will dissolve)

- store at -20˚C

2mM dNTPs for PCR

• the original concentration for each of the dNTPs (dCTP, dTTP, dATP, & dGTP) is 100mM

• add 20µl of each (total 80µl) to 920µl of ddH2O to get 1ml of stock 2mM solution

• dispense into aliquots and freeze at -20˚C