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Crystalgen Inc.
1-800-216-3287 Product Profile
 

Instruction for Ten-Minute DNA Release Kit
The DNA kit is designed to obtain DNA extract from mice tail, ear or toe for PCR within 10-30
minutes.
It is a rapid screening method for transgenic mice.
Catalog Number DNA Release Buffer Samples Storage & Use
Name Volume
100
-Can be used for 2 years at 20 °C.
-or 4°C for 6 months
-Warm to RT before using if it was kept
below 4°C.
300-907-004
Buffer A 20 ml
Buffer B 4 ml
Tools
• Thermo-machine or water bath which can be set for a constant temperature between 55° -
95°C;
• Micro Centrifuge;
• 1.5ml micro tubes, pipette tips, and pipette with 10-200ul scale.

Protocol
Steps Action Example
Step #1 • Place 1.5-3.0 mm section of a mouse tail or 2 x3 mm of its ear or toe
tissue (or any other tissue** or material stained with body fluid**) in
a micro tube. Add 200ul of the Buffer A to the tube. Heat at 93 °C for
3 Min. Cool down 2 Min, Add 40ul Buffer B to the tube.


Step #2 • Put the tube in a thermomixer machine or water bath at 55° C, shaking
at 200-400rpm for 10-30 minutes (If shaking is not available extend
the time to 30 minutes*) to release the DNA.
* For older mouse tail , you will get a better result in 20-30 minutes
Step #3 • Flick the tube 3-4 times, heat the tube at 95° C for 5 minutes.
Step #4 • Centrifuge at 10,000 rpm for 5 min at room temperature.
Notes
You may see the part of tail is still in the solution, but the DNA has
already been released and it is ready for PCR.
• Take the 1-2ul DNA extract as the template to run PCR in 20-25 ul
reaction volume.
• Transfer the rest of DNA extract to a fresh tube at 4 °C. The DNA can
be used for 2-4 weeks, or to be kept at -20 °C for long time storage.
• Usually, the kit works well for 100-1200 bp PCR fragment.
• If your PCR fragment is over 1200 bp, or if you want to get high yield
of the DNA to do Southern et al, you should use the Ten-Minute DNA
Release Kit-2 to obtain the purified DNA within 1 hour. The purified
DNA can be used in both the Southern and PCR.


Crystalgen Inc. Product Profile
**Using Different Organs’ Tissue or Materials Stained by Body Fluid
1. Cut 1-3mm3 (1-12 mg) of the sample;
2. If the sample is clean (ex. clear skin tissue), follow the protocol;
3. If the sample is stained with blood or any other dirty trace:
a. rinse it 2-3 times with PBS without Ca++ and Mg++;
b. dry the tissue on a paper towel; then, to follow the work table.
Ordering information
Name Cat. # Price/Kit Reactions
Ten-Minute DNA Extraction Kit
300-907-004
ask/Kit 100
ask/10 Kits 10 x 100
 


 

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Last modified: 09/28/09