Geneflow
Q-Spin Gel Extraction/PCR Purification Kit
·
Up to 25 µg 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
µl
DNA
fragments cut from agarose preparative gel after electrophoresis of
DNA ladder and purified using Q-Spin Gel/PCR DNA Purification Kit (5
µl/lane). DNA was eluted from spin column with 40 µl 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 µg 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
Ordering Information
| Order Code |
Unit |
Description |
Price |
|
K1-0030 |
Kit |
Geneflow Q-Spin Gel Extraction/PCR Purification Kit, 50 Preps |
£38.5 |
|
K1-0040 |
Kit |
Geneflow Q-Spin Gel Extraction/PCR Purification Kit, 250 Preps |
£155 |
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 µl 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 µl 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 µl 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 50-60°C before use. |
|
Gel is difficult to
dissolve
|
Add more Binding Buffer
and repeat solubilisation step 2. |
For research use
only.
March 2006
