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Team:TU Darmstadt/Notebook/Materials/PCR buffers and mastermixtures
From 2014.igem.org
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| - | <!--TYPO3SEARCH_begin--><div id="c103" class="csc-default"><div class="csc-header csc-header-n1"><h1 class="csc-firstHeader">10x Taq buffer</h1></div><div class=""><p>100 ml batch</p></div><div class=""></div><p>- 500 mM KCl<span style="white-space:pre"> | + | <!--TYPO3SEARCH_begin--><div id="c103" class="csc-default"><div class="csc-header csc-header-n1"><h1 class="csc-firstHeader">10x Taq buffer</h1></div><div class=""><p>100 ml batch</p></div><div class=""></div><p>- 500 mM KCl<span style="white-space:pre"> </p><div class=""><p>- 0,8 % Triton X-100<span style="white-space:pre"> </span> 800 µL</p></div><div class=""><p>- 100 mM Tris-HCl (pH 8.8 bei 25°C)<span style="white-space:pre"> </span>up to 100 ml</p></div><div class=""></div><div class=""><p>Autoclaving/ sterile filtration after mixing.</p></div></div><div id="c104" class="csc-default"><div class="csc-header csc-header-n2"><h1>10x Pfu buffer</h1></div><div><p>100 ml batch</p></div><div></div><div><p>- 200 mM Tris-HCl (pH 8.8 bei 25°C)<span style="white-space:pre"> </span>2.42 g</p></div><div><p>- 100 mM (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub><span style="white-space:pre"> </span> 1.32 g</p></div><div><p>- 100 mM KCl<span style="white-space:pre"> </span> 0.75 g</p></div><div><p>- 20 mM MgSO<sub>4</sub> x 7 H<sub>2</sub>O<span style="white-space:pre"> </span>0.49 g</p></div><div><p>- 1 % Triton X-100<span style="white-space:pre"> </span> 1 ml</p></div><div><p>- ddH<sub>2</sub>O<span style="white-space:pre"> </span> up to 100 ml</p></div><div></div><div><p>+ 1 ml 100 mg/ml BSA (-> end conc. 1 mg/ml)</p></div><div></div><div><p>Autoclaving/ sterile filtration after mixing.</p></div></div><div id="c105" class="csc-default"><div class="csc-header csc-header-n3"><h1>2x Taq Mastermixture</h1></div><div><p>1250 µL batch</p></div><div></div><div><p>- 250 µL 10x Taq buffer</p></div><div><p>- 25 µL Taq polymerase</p></div><div><p>- 50 µL 50mM MgCl<sub>2</sub></p></div><div><p>- 50 µL 10 mM dNTPs</p></div><div><p>- 75 µL DMSO</p></div><div><p>- 800 µL ddH<sub>2</sub>O</p></div></div><div id="c106" class="csc-default"><div class="csc-header csc-header-n4"><h1>9:25 Pfu Mastermixture </h1></div><div><p>450 µL batch</p></div><div></div><div><p>- 250 µL 10x Pfu buffer BSA</p></div><div><p>- 50 µL Pfu polymerase</p></div><div><p>- 50 µL dNTPs (10 mM)</p></div><div><p>- 50 µL MgCl2 (50 mM)</p></div><div><p>- 50 µL DMSO</p></div></div><!--TYPO3SEARCH_end--> |
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Revision as of 17:51, 17 October 2014
10x Taq buffer
100 ml batch
- 500 mM KCl
- 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
