Contents of the Kit
|Gel lysis buffer ||:65 ml x 1 bottle || Precipitation solution ||:65 ml x 1 bottle |
|Co-precipitation solution ||:0.5 ml x 1 tube || || |
Storage Condition: 0-5ºC
Shipping Condition: ambient temperature
Required Equipment and Materials
microcentrifuge, transiluminater or UV lamp, vortex mixer, 100 μl and 1000 μl adjustable pipettes, ethanol
Get pureDNA Kit-Agarose enables isolation and purification of double-stranded DNA from agarose gel in three easy steps: 1) lysis of agarose gel, 2) removal of agarose gel, and 3) DNA recovery using ethanol precipitation. Double-stranded DNA can be isolated from up to 200 mg of agarose gel using 1.5 ml tubes. To isolate DNA from a larger quantity of agarose gel, simply increase the volume of each solution. This kit does not require phenol, chloroform, centrifugal columns, or filtration tubes. The extracted double-stranded DNA can be utilized for restriction enzyme digestion, ligation, PCR, and other enzymatic reactions.
♦ No or low DNA recovery
a) Completely dissolve the agarose gel slice by vortexing in step 2. It may be difficult to see undissolved agarose gel pieces in Gel lysis buffer.
b) Make sure there is a DNA pellet at the bottom of the tube before discarding the supernatant in steps 8 and 9.
♦ Difficulty dissolving the agarose gel slice in step 3
a) Crush the gel slice in a 1.5 ml tube with a pipette tip.
b) Vortex the tube every 2 minutes during solubilization of the gel slice at 60ºC.
♦ Too much precipitate after centrifugation in step 5
a) Completely dissolve the agarose gel slice before adding precipitation solution.
b) Thoroughly mix the solution by vortexing after adding Precipitation solution.
c) Increase the centrifugation time if 12,000-14,000 rpm (10,000 g) is difficult to achieve.
♦ Low DNA purity
a) Do not disturb the white precipitate in step 6.
b) Remove as much of the supernatant as possible in steps 8 and 9.
♦ Degradation of isolated DNA
a) Minimize the UV irradiation time during preparation of agarose gel slice in step 1.
b) Avoid using short wavelength UV light.