Editing Taq Buffer

== NEB Taq Buffer, 10 X ==

<span>100 mM Tris-HCl, 500 mM KCl, and 15 mM MgCl<sub>2</sub>, pH 8.3 @ 25°C.</span>

'''1'''. 100 ml of 1M Tris-HCl, pH 8.30<br/>'''Trizma HCl: '''6.14 g<br/>'''Trizma Base: '''7.40 g

ultrapure water to 100 ml

<span>'''2'''. 50 ml of 150 mM MgCl<sub>2</sub> · 6H<sub>2</sub>O: MW 203.3</span>

<span>&nbsp;1.53 g, add ultrapure water to 50 ml</span>

<span>'''3'''. 1% Tween 20:&nbsp; 5 g in 50 ml ultrapure water.</span>

<span>'''To prepare 50 ml 10 X Buffer without Tween 20:'''</span>

<span>5 ml 1M Tris-HCl, pH 8.30.</span>

<span>5 ml 150 mM <span>MgCl<sub>2</sub></span></span>

<span>0.05 * 0.5 * 74.55=1.86 g KCl.</span>

<span>Ultrapure Water to 50 ml.</span>

<span>'''To prepare 50 ml 10 X Buffer without Tween 20:'''</span>

<span>5 ml 1M Tris-HCl, pH 8.30.</span>

<span>5 ml 150 mM <span>MgCl<sub>2</sub></span></span>

<span>+ 0.05 * 0.5 * 74.55=1.86 g KCl.</span>

<span>+&nbsp; 0.5 ml of 10% Tween 20</span>

<span>ultrapure Water to 50 ml.</span>

<span>'''1 M Trehalose MW 342.29'''</span>

<span>17 g in 50 ml ultrapure water.</span>



== <span>10X Taq Buffer with KCl</span> ==

<span>100 mM Tris-HCl (pH 8.8 or 8.3 at 25°C), 500 mM KCl, 15 mM MgCl2, 0.8% (v/v) Nonidet P40 or Triton X-100.</span>

== <span>10X Taq Buffer with (NH4)2SO4</span> ==

<span>750 mM Tris-HCl (pH 8.8 at 25°C), 15 mM MgCl2, 200 mM (NH4)2SO4, 0.1% (v/v) Tween 20.</span>



<span>2X Taq MasterMix</span>

<span>For StepOne ABI</span>

{| style="width: 500px" cellspacing="1" cellpadding="1" border="1"
|-
| <span>Components</span><br/>
| <span>ul</span><br/>
|-
| <span>H<sub>2</sub>O</span><br/>
| <span>15.15</span><br/>
|-
| <span>Buffer 10 X</span><br/>
| <span>2.5</span><br/>
|-
| <span>MgCl<sub>2,</sub> 50 mM</span><br/>
| <span>1.5</span><br/>
|-
| <span>dNTPs 10 mM</span><br/>
| <span>0.5</span><br/>
|-
| <span>Primers 5 uM</span><br/>
| <span>2</span><br/>
|-
| <span>BSA 10 mg/ml</span><br/>
| <span>0.25</span><br/>
|-
| <span>Taq&nbsp; 5 U/ul</span><br/>
| <span>0.2</span><br/>
|-
| <span>Sybr (10x)</span><br/>
| <span>0.4</span><br/>
|-
| <span>Rox (5x)</span><br/>
| <span>0.5</span><br/>
|-
| <span>cDNA</span><br/>
| <span>2</span><br/>
|}

<span>Increase SYBR and decrease ROX concentrations for Stratagene Mx3005P and BioRad iCycler</span>

<span>Sigma Jump Start Taq Ready Mix.<br/>5ul SYBR green mix<br/>MgCl2 1.2 ul<br/>0.4 ul forward primer (10 pmol)<br/>0.4 ul reverse primer (10 pmol)<br/>1 ul water<br/>2ul cDNA (1/4 diluted cDNA)</span>

<span>For High Resolution Melting: use&nbsp;Biotium EvaGreen&nbsp; and Promega BRYT Green, becuase SYBR Green&nbsp; inhibits PCR at higher concentrations. EvaGreen at high enough concentrations to saturate the double strand DNA</span>

== <span>Sigma AccuTaq LA and REDAccuTaq LA Buffer</span> ==

<span>AccuTaq LA: 500 mM Tris-HCl, adjust pH to 9.3 using NH4OH, 150 mM (NH4)2SO4, 25 mM MgCl2, 1% Tween 20</span>

<span>REDAccuTaq LA </span>: <span>500 mM Tris-HCl, adjust pH to 9.3 using KOH, 150 mM (NH4)2SO4, 25 mM MgCl2, 1% Tween 20</span>
[[Category:Buffer Recipe]]