PCR Kleen™ Spin Purification Module

Image
pcr_kleen_spin.jpg

Overview

The PCR Kleen Spin Purification module uses size exclusion chromatography to remove unincorporated primers, nucleotides, salts, and enzymes from PCR products.

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Description

Description

The PCR Kleen Spin purification module uses size exclusion chromatography to remove unincorporated primers, nucleotides, salts, and enzymes from PCR products. This protocol purifies PCR products using simple spin columns that remove the contaminating molecules by absorbing them into porous beads. Larger molecules, such as PCR products greater than 200 bp, cannot enter the beads and are eluted from the column into a collection tube.

Following the purification step, both the unpurified and the purified PCR products can be resolved side-by-side using agarose gel electrophoresis and stained to visually assess the removal of oligonucleotide primers. Once purified, the PCR product is ligated into the pJet1.2 plasmid vector.

The PCR Kleen Spin purification module is an integral component of the Cloning and Sequencing Explorer Series, where it is used to purify a portion of GAPDH gene that has been amplified. The module can also be purchased separately and used in other experiments to:

  • clean any PCR reaction prior to ligation
  • remove primers prior to subsequent nested PCR reactions
  • remove nucleotides before direct DNA sequencing

Specifications

Specifications

Method
Size exclusion chromatography in spin column
Applications
Removal of nucleic acids <32 bases and other reaction mixture components from PCR products >200 bp
Capacity
25–100 µl
Equilibration buffer
10 mm Tris, 1mM EDTA, pH 7.4
Sample preparation time
<4 min
Yield
50–80% recovery

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7326300EDU
Fourth module in Cloning and Sequencing Explorer Series classroom study kit, includes PCR Kleen Spin columns, test tubes, curriculum, for 12 workstations; education use only

Documents

Documents

Number Description Options
4006142
Instruction Manual, Quantum Prep PCR Kleen Spin Columns, Biotechnology Explorer, Rev B
2311
PCR Kleen Spin Columns Product Information Sheet, Rev D
2950
Comparison of PCR Kleen Spin Columns to Traditional Methods for Purification of PCR Products Prior to Sequencing, Rev A