Team:TU Darmstadt/Notebook/Materials/PCR buffers and mastermixtures

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Revision as of 16:57, 17 October 2014

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10x Taq buffer

100 ml batch

- 500 mM KCl 3,7 g

- 0,8 % Triton X-100         800 µL

- 100 mM Tris-HCl (pH 8.8 bei 25°C) up to 100 ml

Autoclaving/ sterile filtration after mixing.

10x Pfu buffer

100 ml batch

- 200 mM Tris-HCl (pH 8.8 bei 25°C) 2.42 g

- 100 mM (NH4)2SO4         1.32 g

- 100 mM KCl         0.75 g

- 20 mM MgSO4 x 7 H2O 0.49 g

- 1 % Triton X-100         1 ml

- ddH2O         up to 100 ml

+ 1 ml 100 mg/ml BSA (-> end conc. 1 mg/ml)

Autoclaving/ sterile filtration after mixing.

2x Taq Mastermixture

1250 µL batch

- 250 µL 10x Taq buffer

- 25 µL Taq polymerase

- 50 µL 50mM MgCl2

- 50 µL 10 mM dNTPs

- 75 µL DMSO

- 800 µL ddH2O

9:25 Pfu Mastermixture

450 µL batch

- 250 µL 10x Pfu buffer BSA

- 50 µL Pfu polymerase

- 50 µL dNTPs (10 mM)

- 50 µL MgCl2 (50 mM)

- 50 µL DMSO