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Team:TU Darmstadt/Notebook/Materials/PCR buffers and mastermixtures
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Revision as of 16:19, 17 October 2014
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
