SRMP Lab Skills: Molecular Biology Curriculum - American Museum ...
SRMP Lab Skills: Molecular Biology Curriculum - American Museum ...
SRMP Lab Skills: Molecular Biology Curriculum - American Museum ...
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Science Research Mentoring Program<br />
MOLECULAR BIOLOGY<br />
Session Three: DNA Extraction: WORKSHEET<br />
PCR Reaction<br />
COMPONENT / STOCK SOLUTION<br />
VOLUME (FILL IN)<br />
FINAL CONCENTRATION<br />
10X PCR Buffer, minus Mg<br />
1X<br />
10 mM dNTP mixture (dATP,<br />
dCTP, dTTP, dGTP)<br />
0.2 mM<br />
50 mM MgCl 2<br />
1.5 mM<br />
Primers (20 μM)<br />
0.4 μM each primer<br />
Template DNA (100 ng/μl)<br />
2 ng/μl<br />
Taq DNA Polymerase (5 units/μl)<br />
1.0 unit in reaction<br />
Water<br />
NA<br />
1. What information do you need before you can calculate the volume of each component<br />
2. You will run your PCR product on an agarose gel. To make a 2% agarose gel (w/v), how<br />
much agarose would you put into 50 mL of 1X TAE buffer<br />
3. If your stock solution of TAE is 50X, how much stock solution would you dilute in how<br />
much water to make the 50 mL of 1X buffer<br />
4. TAE buffer (1X, working dilution) is: 40 mM Tris base (short for 2-amino-2-hydroxymethylpropane-1,3-diol),<br />
1.14% acetic acid (v/v), 1 mM EDTA. Calculate how much Tris, acetic acid,<br />
and EDTA stock solution you would need to make 1 L of 50X TAE. The molecular weight of<br />
Tris is 121, and your stock solution of EDTA is 0.5 M.<br />
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