Geneflow Q-Spin Gel Extraction/PCR Purification Kit

  • Up to 25 g of DNA can be purified with a single prep
  • Purifies DNA from agarose gels and PCR mixes
  • DNA fragments 100 bp-10 kb
  • Spin columns with elution volume 25-50 l

DNA fragments cut from agarose preparative gel after electrophoresis of DNA ladder and purified using Q-Spin Gel/PCR DNA Purification Kit (5 l/lane). DNA was eluted from spin column with 40 l of elution buffer.

The Geneflow Q-Spin Gel/PCR kit provides a simple and efficient method for purification of DNA fragments from agarose gel and from PCR mixes. DNA fragments selectively absorbed in the spin column after incubation at 60oC for gel extraction or at room temperature for PCR product with equal volume of binding buffer.

Up to 25 g of DNA can be purified with a single prep using Q-Spin Gel/PCR DNA Purification Kit.

Applications

  • DNA extraction and purification from agarose gel 
  • PCR DNA clean-up

Features

  • Spin column format
  • Equal volume of binding buffer used for agarose solubilisation
  • DNA fragments 100 bp – 10 kb
  • Reproducible yield up to 25 mg of plasmid DNA per prep
  • 12 samples in just 15 minutes
  • Elution volume 25-50 ml

Reagents supplied in kit 

Spin columns, 2-ml collection tube, Binding buffer, Wash solution concentrate,

Elution buffer 5 mM Tris-Cl, pH 8.0

Geneflow Q-Spin Gel Extraction/PCR Purification Kit

The Kit is designed for purification of DNA up to 10 Kb from agarose gels or reaction solutions

(PCRs or restriction digests).

Components    
Spin Columns 50 250
2-ml Collection Tubes 50 250
Binding Buffer 30ml 3 x 50ml
Wash Solution I, concentrate 8ml 5 x 8ml
Elution Buffer, 5 mM Tris-Cl, pH 8.0 5ml 25ml

Notes before use:

1.       Add 50 ml of ethanol (96-100%) to 8 ml of Wash Solution I concentrate.

2.       TE buffer is not recommended for elution.

3.       Bench top micro centrifuge (7,000-12,000 rpm) can be used for all following procedures.

Protocol

 1. For DNA extraction from the gel: Excise the DNA fragment from the agarose gel and transfer to a clean microfuge tube. Weight the gel slice and add equal volume of Binding Buffer, e.g 100 l of the Buffer per 100 mg of gel slice. Excess of the Binding Buffer is preferable. For concentration of agarose >1.5% add twice the volume of the Binding Buffer.

2. Incubate with occasional vortexing for 5-10 min in a water bath (50-65 C) or until the gel is completely dissolved.

3. For DNA purification from reaction mixes (PCR or digests): Add equal volume of Binding Buffer, vortex or mix with pipetting. Incubate at RT for 1 min.

4. Add the above mixtures to the spin column and let to stand at RT for 2 min.

5. Centrifuge for 1 min and discard the flow-through in the tube.

6. Add 500 l of Wash Solution I and spin for 15 seconds. Discard the flow-through solution.

7. Repeat step 6.

8. Centrifuge for 1 minute to remove residual Wash Solution I.

9. Transfer the column into 1.5 ml microfuge tube, cut the tube’s cap with scissors if necessary.  Add 30-50 l of Elution Buffer on to the center part of membrane of the column and incubate for 1- 2 min at room temperature. Spin for 1 min.

10. Transfer the plasmid DNA into a clean tube and store at –20 C.

Troubleshooting

Low DNA yield

 

DNA fragments smaller than 100 bp and bigger than 10 kb may lead to lower recovery of DNA.

Increase the incubation time with Elution Buffer.

Preheat column and Elution Buffer at 50-60C before use.

Gel is difficult to dissolve

 

Add more Binding Buffer and repeat solubilisation step 2.

For research use only.    


 
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