Team:HKUST/Protocols/PCR cleanup

From 2009.igem.org

(Difference between revisions)
Line 69: Line 69:
   h.Centrifuge at 14,000 rpm for 1 min to elute the DNA.<br>
   h.Centrifuge at 14,000 rpm for 1 min to elute the DNA.<br>
<p> Tips: </p>
<p> Tips: </p>
-
In step f, for effective elution, make sure that the elution solution is dispensed onto the membrane center and is absorbed completely.
+
In step f, for effective elution, make sure that the elution solution is dispensed onto the membrane center and is absorbed completely.</p>

Revision as of 15:48, 19 October 2009

Salt and Soap template

a

PCR product clean-up

Purpose: To clean up primers, dNTPs, enzymes, short-failed PCR products in the PCR reaction.

Materials: FAPC Buffer,Wash Buffer, Elution Buffer, PCR Product (all provided in FavorPrepTM PCR Clean-Up Kit)

Procedure:

a.Transfer 30μL of PCR product and add 5 volumes of FAPC buffer to a 1.5mL microcentrifuge tube, then mix well by vortexing.
b.Place a FAPC Column into a Collection Tube and transfer the sample mixture to FAPC Column.
c.Centrifuge at 6,000 rpm for1 min then discard the flow-through.
d.Add 750 μl of Wash Buffer (ethanol added) to FAPC Column. Centrifuge at 6,000 rpm for 1 min, then discard the flow-through.
e.Centrifuge at 14,000 rpm for an additional 3 min to dry the column.
f.Place FAPC Column into an Elution Tube.
g.Add 40 μl of Elution Buffer or ddH2O (pH 7.0~8.5) to the membrane center of FAPC Column. Stand FAPC Column for 2 minutes.
h.Centrifuge at 14,000 rpm for 1 min to elute the DNA.

Tips:

In step f, for effective elution, make sure that the elution solution is dispensed onto the membrane center and is absorbed completely.

HKUST