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

From 2014.igem.org

(Difference between revisions)
 
(9 intermediate revisions not shown)
Line 4: Line 4:
<html>
<html>
-
<div id="contentWrap" class="container_24">
+
 
-
<div id="breadcrumbs" class="grid_24">
+
-
<p>Sie sind hier:&nbsp; <a href="https://2014.igem.org/Team:TU_Darmstadt/" >wiki</a> &rsaquo;&nbsp;<a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook" >Notebook</a> &rsaquo;&nbsp;<span class="current"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials" >Materials </a> &rsaquo;&nbsp;<span class="current"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/PCR_buffers_and_mastermixtures"> PCR buffers and mastermixtures </a></span></p>
+
-
</div>
+
<div id="leftNavi" class="grid_5">
<div id="leftNavi" class="grid_5">
<nav>
<nav>
-
<ul class="menu"><li class="first"><a href="https://2014.igem.org/Team:TU_Darmstadt/" >Home</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Project" >Project</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results" >Results</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/PolicyandPractices" >Policy & Practices</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Achievements" >Achievements</a></li><li class="active"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook" >Notebook</a><ul class="menu2"><li class="first"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal" >Labjournal</a></li><li class="active"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials" >Materials</a><ul class="menu3"><li class="first"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Strains" >Strains</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Media" >Media</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Enzymes" >Enzymes</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Buffers" >Buffers</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Stock_Solution" >Stock Solution</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Primers" >Primers</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Equipment" >Equipment</a></li><li class="last active"><a href="https://2014.igem.org/Team:TU_Darmstadt/Materials/PCR_buffers_and_mastermixtures" >PCR buffers and mastermixtures</a></li></ul></li><li class="last"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Methods" >Methods</a></li></ul></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Team" >Team</a></li><li class="last"><a href="https://2014.igem.org/Team:TU_Darmstadt/Gallery" >Gallery</a></li></ul>
+
<ul class="menu"><li class="first"><a href="https://2014.igem.org/Team:TU_Darmstadt" >Home</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Project" >Project</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results" >Results</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/PolicyandPractices" >Policy & Practices</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Achievements" >Achievements</a></li><li class="active"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook" >Notebook</a><ul class="menu2"><li class="first"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal" >Labjournal</a></li><li class="active"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials" >Materials</a><ul class="menu3"><li class="first"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Strains" >Strains</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Media" >Media</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Enzymes" >Enzymes</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Buffers" >Buffers</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Stock_Solution" >Stock Solution</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Primers" >Primers</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials/Equipment" >Equipment</a></li><li class="last active"><a href="https://2014.igem.org/Team:TU_Darmstadt/Materials/PCR_buffers_and_mastermixtures" >PCR buffers and mastermixtures</a></li></ul></li><li class="last"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Methods" >Methods</a></li></ul></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Team" >Team</a></li><li class="last"><a href="https://2014.igem.org/Team:TU_Darmstadt/Gallery" >Gallery</a></li></ul>
</nav>
</nav>
</div>
</div>
Line 18: Line 16:
<div id="wikicontent" class="grid_19">
<div id="wikicontent" class="grid_19">
-
<!--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">         </span>3,7 g</p><div class=""><p>- 0,8 % Triton X-100<span style="white-space:pre"> </span>&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;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>&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;1.32 g</p></div><div><p>- 100 mM KCl<span style="white-space:pre"> </span>&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;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>&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;1 ml</p></div><div><p>- ddH<sub>2</sub>O<span style="white-space:pre"> </span>&nbsp; &nbsp; &nbsp; &nbsp; up to 100 ml</p></div><div></div><div><p>+ 1 ml 100 mg/ml BSA (-&gt; 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-->
+
<!--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"> </p></div><div class=""><p>- 100 mM Tris-HCl (pH 8.8 bei 25°C)<span style="white-space:pre"> </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"> </p></div><div><p>- 100 mM (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub><span style="white-space:pre"> </span>&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;</p></div><div><p>- 100 mM KCl<span style="white-space:pre"> </span>&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;</p></div><div><p>- 20 mM MgSO<sub>4</sub> x 7 H<sub>2</sub>O<span style="white-space:pre"> </span></p></div><div><p>- 1 % Triton X-100<span style="white-space:pre"> </span>&nbsp; &nbsp; &nbsp; &nbsp;&nbsp;</p></div><div><p>- ddH<sub>2</sub>O<span style="white-space:pre"> </span>&nbsp; &nbsp; &nbsp; &nbsp;</p></div><div></div><div><p>- 1 ml 100 mg/ml BSA (-&gt; 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-->
</div>
</div>
</html>
</html>

Latest revision as of 18:09, 17 October 2014

Home


10x Taq buffer

100 ml batch

- 500 mM KCl

- 0,8 % Triton X-100

- 100 mM Tris-HCl (pH 8.8 bei 25°C)

Autoclaving/ sterile filtration after mixing.

10x Pfu buffer

100 ml batch

- 200 mM Tris-HCl (pH 8.8 bei 25°C)

- 100 mM (NH4)2SO4         

- 100 mM KCl         

- 20 mM MgSO4 x 7 H2O

- 1 % Triton X-100         

- ddH2O        

- 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