Editing Phusion Polymerase (section)

== Pfu Buffer ==

10 mM KCl

10 mM (NH4)2SO4

20 mM Tris-HCl, pH 8.8

2 mM MgSO4

0.1% Triton X-100

100 ug/ml BSA

'''Herculase PCR Buffer'''

50 mM Tricine, pH 9.1

8 mM (NH4)2SO4

2.3 mM MgCl2

0.1% Tween-20

75 ug/ml BSA

For PCR, add 2 U of Pfu dUTPase, 5 U Pfu/Phusion

'''Taq Buffer'''

10 mM Tris-HCl (pH 8.8)

50 mM KCl

2 mM MgCl2

0.1% Triton&nbsp;X-100



'''Buffer from websites'''

<span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">1. 10X Taq Buffer with KCl</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">100mM Tris-HCl (pH 8.8 at 25 °C),</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">500mM KCl,</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">0.8% Nonidet P40.</span><br/><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">2. 10X Taq Buffer with KCl and 15mM MgCl2</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">100mM Tris-HCl (pH 8.8 at 25 °C),</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">500mM KCl,</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">0.8% Nonidet P40,</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">15mM MgCl2.</span><br/><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">3. 10X Taq Buffers with KCl Set</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">100mM Tris-HCl (pH 8.8 at 25 °C),</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">500mM KCl,</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">0.8% Nonidet P40,</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">MgCl2 different in each buffer (12.5-30mM)</span><br/><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">4. 10X Taq Buffer with (NH4)2SO4</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">750mM Tris-HCl (pH 8.8 at 25 °C),</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">200mM (NH4)2SO4,</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">0.1% Tween 20.</span><br/><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">5. 10X Taq Buffer with (NH4)2SO4 and 20mM MgCl2</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">750mM Tris-HCl (pH 8.8 at 25 °C),</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">200mM (NH4)2SO4,</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">0.1% Tween 20,</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">20mM MgCl2.</span><br/><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">6. 10X Taq Buffer without Detergent</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">100mM Tris-HCl (pH 8.8 at 25 °C),</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">500mM KCl.</span><br/><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">7. 10X Pfu Buffer with 20mM MgSO4</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">200mM Tris-HCl (pH 8.8 at 25 °C), 100mM (NH4)2SO4, 100mM KCl, 1% (v/v) Triton X-100, 1mg/ml BSA and 20mM MgSO4.</span><br/><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">8. 10X Pfu Buffer</span><br/><span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;">200mM Tris-HCl (pH 8.8 at 25 °C), 100mM (NH4)2SO4, 100mM KCl, 1% (v/v) Triton X-100, 1mg/ml BSA.</span><span style="font-family: Arial, Helvetica, sans-serif; font-size: 14px;"></span>

<span style="color: rgb(0, 0, 0); font-family: arial, sans-serif; font-size: 15.2px;"></span><span style="font-family: Arial, Helvetica, sans-serif; font-size: 14px;"></span>



References

1. Wang, Y., Prosen, D. E., Mei, L., Sullivan, J. C., Finney, M., Vander Horn, P. B. (2004) A novel strategy to engineer DNA polymerases for enhanced processivity and improved performance in vitro.&nbsp;Nucleic Acids Res.&nbsp;32: 1197–1207.&nbsp;«PubMedCentral»