Instructions for Isohelix DNA Isolation kit: DDK-50 / DDK-3

Instructions for Isohelix DNA Isolation kit: DDK-50 / DDK-3
Product Details
Isohelix Buccal DNA Isolation Kits have been specifically formulated to produce high DNA yield and purity from buccal
swabs. The kits have been fully optimised at Cell Projects for use on buccal cell samples and offer reduced handling times,
increased DNA yields and many other important technical benefits for their use in manual, 96-well or other high
throughput formats.
Key Benefits
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Optimised for buccal cells
Fast handling times
High purity and yield
No solvent based chemicals
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Protocol integrated to swabs
Manual or high throughput formats
No columns or filtration
Less consumables wastage
Kit Contents
Isohelix DDK DNA Isolation Kits
Catalogue No.
Number of preps
Solution LS (Lysis buffer)
Proteinase K
Solution CT (Capture buffer)
Solution TE (Re-hydration buffer)
DDK-50
50
DDK-3
3
25ml
2 x 11mg*1
25ml
15ml
1.5ml
2.2mg*2
1.5ml
1.0ml
Storage temperature
Room temperature
40C after reconstitution
Room temperature
Room temperature
*1 Reconstitute each vial with 550µl sterile ddH2O before first use, store at 40C after reconstitution.
*2 Reconstitute vial with 110µl sterile ddH2O before first use, store at 40C after reconstitution.
Storage
Isohelix DDK DNA Isolation Kits are shipped at ambient temperature.
Please note that on arrival the kit components should be stored according to the table above.
The kits are stable up to the expiry date if stored as instructed. See box label for expiry date.
Equipment and reagents to be supplied by user
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Water bath or heating block at 600C
Pipettes with disposable tips
Microcentrifuge (with rotor for 2 ml tubes)
1.5ml microcentrifuge tubes
Vortexer
Before Starting
1.
2.
3.
Prepare a waterbath at 600C
If a precipitate has formed in solution LS, warm at 370C for a few minutes
Reconstitute the Proteinase K by adding the appropriate amount of sterile ddH2O as shown above.
Technical Assistance
If you have any questions regarding the use of this kit or other Isohelix products please contact us by email at info@isohelix
.com or for further information visit the website at www.isohelix.com
Safety and Use of the DDK DNA Isolation kits
The DDK kits are intended for use by qualified professionals trained in potential laboratory hazards and
good laboratory practise. If direct information is not available on any of our compounds this should not be
interpreted as an indication of product safety.
This kit has been designed for research use only
Version May 2015
Isohelix is a division of Cell Projects
For swab or DNA isolation queries email: info@isohelix.com www.isohelix.com
Molecular Biology Solutions www.cellprojects.com
Page 1 of 2
DNA Isolation Protocol for DDK-3/DDK-50
Part A – DNA Stabilisation
1.
Add 500ul LS solution to the tube containing the buccal swab.
2.
Add 20µl PK solution to the tube containing the buccal swab and LS solution. Vortex briefly.
At this point the DNA is stabilised. You may continue with the DNA isolation or store the stabilised swab in a
sealed tube at room temperature for at least 3 ½ years.
Part B – DNA Isolation
3.
Place the tube containing the swab, LS solution and PK solution in a 600C water bath for 1 hour. Vortex briefly.
4.
Transfer the liquid in the tube (approx. 400µl) into a 1.5ml centrifuge tube using a sterile pipette tip.
5.
Optional step to increase yield:
If using SK-1 swabs, tip the swab head into a sterile 1.5ml centrifuge tube so that the swab head is uppermost.
Spin the tube briefly and using a sterile pipette tip add the recovered supernatant to the 400µl collected
previously.
If using SK-2 swabs, spin the SK-2 tube containing the swab head briefly and using a sterile pipette tip add the
recovered supernatant to the 400µl collected previously
6.
Add 400µl CT solution to the tube, (500µl if using the optional step 5). Vortex briefly.
7.
Place the tube in a microcentrifuge (with hinge positioned outwards so the liquid can be removed from the
opposite side) and spin at maximum speed (13.4K rpm/12,000 x g) for 7 minutes to pellet the DNA. Note the
pellet may not be visible.
8.
Remove all the supernatant carefully with a pipette tip taking care not to disturb the DNA pellet.
9.
Re-spin the tube briefly and remove any remaining liquid. Note it is important to remove all of the liquid.
10. Add 150µl TE solution to the tube. This volume may be decreased to as little as 30µl if a higher concentration of
DNA is required.
11. Leave for at least 5 minutes at room temperature for the DNA to re-hydrate, longer if a reduced volume of TE has
been used. Vortex briefly.
12. Re-spin the tube for 15 minutes at maximum speed (13.4K rpm/12,000 x g) to remove undissolved debris.
Transfer the supernatant to a sterile 1.5ml tube, being careful not to disturb the pellet.
The DNA sample is now ready for use in downstream applications such as amplification.
Store the DNA sample at 40C for short term storage or -200C for long term storage.
The expected yield from a buccal swab is on average 1 to 10µg DNA (5 to 70ng/µl) from an adult.
Our DQC-50 DNA Quality Check Kit is designed specifically to confirm presence of and to test both the quality and quantity
of your human DNA by a quick PCR test before you start downstream testing.
Other Isohelix Products
Isohelix GeneFixTM Saliva DNA collection and purification kits
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GeneFixTM Saliva DNA Collectors for collecting, storing, stabilising and transporting 2ml saliva samples: GFX-02
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GeneFixTM Saliva DNA Mini Kit for isolation of high purity DNA from GeneFix TM Saliva samples: GSS-50/GSS-5
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GeneFixTM Saliva DNA Midi Kit for isolating high purity DNA from 2ml GeneFixTM Saliva samples: GMS-12/GMS-2
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GeneFixTM Saliva-Prep DNA Kit for isolating DNA from 2ml GeneFixTM Saliva samples: GSP-48/GSP-12/GSP-2
Isohelix DNA Buccal Swabs
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High yields, blood alternative, reproducible, easy to use, different formats for various extraction methodologies.
Isohelix DNA Silica Gel Dri-Capsules
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For use with SK-1 swab kits, air-dries swab in tube giving extended storage times without loss of stability: SGC-50
Isohelix Buccal DNA Isolation and Handling kits
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High yielding DNA Isolation kits for the isolation of DNA from buccal swabs: DDK-50
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Xtreme DNA Kit for isolation of high purity DNA from buccal swabs: XME-50
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DNA Clean-Up kits for additional purification of DNA samples to give ultra-pure DNA: XCU-50
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DNA stabilising kits for the stable storage of DNA at Room Temperature for long periods: DSK-50
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DNA quality check by PCR to confirm quality of DNA prior onward experimentation: DQC-50
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DNA Release kits – quick and easy kit for PCR-ready DNA from buccal swabs in under 20 minutes: BEK-50
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Isohelix Spin+CollectTM sample recovery devices to increase yields from swabs and other materials: SC/ST-100
Version May 2015
Isohelix is a division of Cell Projects
For swab or DNA isolation queries email: info@isohelix.com www.isohelix.com
Molecular Biology Solutions www.cellprojects.com
Page 2 of 2