Team:Toronto/Experiment-PureLink PCR Purification protocol
PureLink ® PCR Purification protocol
Introduction
Minor changes were made to the protocol supplied with the Invitrogen PureLink® PCR Purification Kit
Materials
- Reagents
- PureLink ® Binding Buffer (B2)
- PureLink ® Spin Column
- Wash Buffer (supplied with kit)
- Elution Buffer (10 mM Tris-HCl, pH 8.5)
- Disstilled water >7.0 pH
- Materials
- PureLink ® Spin Column
- Collection Tube
- centrifuge
Procedure
- Protocol
- Add 4 volumes of PureLink ® Binding Buffer (B2) with isopropanol to 1 volume of the PCR product (50–100 μL). Mix well.
- Remove a PureLink ® Spin Column in a Collection Tube from the package.
- Add the sample with the appropriate Binding Buffer (from step 1 of this procedure) to the PureLink ® Spin Column.
- Centrifuge the column at room temperature at 13,000 × g for 1 minute.
- Discard the flow through and place the spin column into the collection tube.
- Add 650 μL of Wash Buffer with ethanol to the column.
- Centrifuge th e column at room temperature at 13,000 × g for 1 minute. Discard the flow through from the collection tube and place the column into the tube.
- Centrifuge the column at maximum speed (15, 000 x g)at room temperature for 2–3 minutes to remove any residual Wash Buffer. Discard the collection tube.
- Place the spin column in a clean 1.7-mL PureLink ® Elution Tube supplied with the kit.
- Add 50 μL of Elution Buffer (10 mM Tris-HCl, pH 8.5) or sterile, distilled water (pH >7.0) to the center of the column.
- Incubate the column at room temperature for 1 minute. Centrifuge the column at maximum speed for 2 minutes.
- The elution tube contains the purified PCR product. Remove and discard the column. Store the purified PCR product at –20°C or use the PCR product for the desired downstream application.
- Reference
- PureLink® PCR Purification protocol. From:http://tools.thermofisher.com/content/sfs/manuals/purelink_pcr_man.pdf
- Changelog
- Adapted from iGEM 2015 website 5/26/2016
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