Rapid Blood Clot DNA Purification Kit

Rapid Blood Clot DNA Purification Kit

product composition

Product storage and expiry date

1．  Proteinase K stock solution and Buffer SP at -20 ℃.

2．  Buffer L7 can be transported at room temperature. After receiving the product, please store it at 2~8℃.

3．  If other reagents and items are stored at normal temperature (0~30℃), their performance will remain unchanged within two years; if the product is stored at 2~8℃, the validity period of the product can be extended to more than two years (2~8℃ Stored products should be allowed to return to room temperature before use).

Product Description

purifying total DNA from 300~400 mg blood clots . There is no need to grind blood clots, just add lysis solution for digestion. The dissolved blood clot is precipitated with hemoglobin in Buffer L7. The DNA in the supernatant is bound to the purification column. The degraded protein and PCR inhibitors are removed by filtration. The DNA on the purification column is washed with Buffer WB and washed with Buffer TE. Once removed, it can be used for various molecular biology experiments.

Reagents and items that users need to prepare by themselves

1．  Anhydrous ethanol

2．  1.5 ml centrifuge tube, pipette and tip (to avoid contamination between samples, please use pipette tips with filter elements)

3．  Disposable gloves and protective equipment and tissues

4．  Desktop small volume centrifuge (can be equipped with a rotor for centrifuging 1.5 ml centrifuge tubes and 2 ml centrifuge tubes)

5．  Water bath and vortex shaker

Preparation before use

1.      If the centrifuge has a refrigeration function, please set the temperature to 25°C.

2.      Set the water bath temperature to 56°C and incubate Buffer PL and Buffer TE to 56°C.

3.      Add absolute ethanol to Buffer WB according to the instructions on the label of the reagent bottle, and check the box on the label to mark "Ethanol has been added".

Steps :

1.       Use a 1.5 ml centrifuge tube to weigh 300~400 mg of blood clot. According to the volume of the blood clot (converted according to 1 mg = 1 µl ), add Buffer PL until the final volume is 500 µl.

* For example, for a blood clot weighing 360 mg, 140 µl Buffer PL should be added.

* If you use a scalpel to remove the blood clot, use the tip of the knife to chop the blood clot into pieces and then transfer it to a 1.5 ml centrifuge tube to speed up the dissolution of the blood clot.

* Do not use more than 400 mg of blood clot, otherwise the digestion capacity of Proteinase K will be exceeded.

2.       Add 20 µl Proteinase K stock solution, then add 15 µl Buffer SP, and vortex for about 15 seconds to mix. 56°C water bath for 10 minutes.

* Vortex for a few seconds every 3 minutes during the water bath to help dissolve the blood clot.

* The blood clot will become viscous after dissolving. If there are still a small amount of undissolved blood clot particles after 10 minutes, it will not affect subsequent operations.

3.       Add 500 µl Buffer L7, cover the tube, shake the centrifuge tube vigorously 3 to 5 times, and then vortex to mix for 30 seconds. Centrifuge at highest speed (≥13000 rpm) for 1 minute.

4.       Pipette the supernatant (about 700 µl) and add it to the nucleic acid purification column (the nucleic acid purification column is placed in a 2 ml centrifuge tube), cover the tube cap, and centrifuge at 12,000 rpm for 30 seconds.

* It is better to aspirate less supernatant than to aspirate the sediment at the bottom of the tube . The sediment will seriously affect the purity of the final DNA.

* If the supernatant is dark in color, you can observe the precipitation by pointing it at the light source.

5.       Discard the filtrate in the 2 ml centrifuge tube, place the nucleic acid purification column back into the 2 ml centrifuge tube, add 800 µl Buffer WB to the nucleic acid purification column, cap the tube, and centrifuge at 12,000 rpm for 30 seconds.

* There is no need to discard the filtrate completely. If you want to avoid contamination of the centrifuge by the filtrate adhering to the mouth of the centrifuge tube, place the 2 ml centrifuge tube upside down on a paper towel and tap it once.

* Confirm that absolute ethanol has been added to Buffer WB.

6.       Discard the filtrate in the 2 ml centrifuge tube, place the nucleic acid purification column back into the 2 ml centrifuge tube, add 300 µl Buffer WB to the nucleic acid purification column, cover the tube, and centrifuge at the highest speed ( ≥ 13000 rpm ) for 1 minute.

7.       Discard the 2 ml centrifuge tube, place the nucleic acid purification column in a clean 1.5 ml centrifuge tube, add 100~200 µl Buffer TE incubated at 56°C in the center of the membrane of the purification column, cover the tube cap, and let stand at room temperature for 1 minute. , centrifuge at 12,000 rpm for 30 seconds.

* Be careful not to let the filtrate touch the bottom of the nucleic acid purification column when taking out the nucleic acid purification column. If the nucleic acid purification column is contaminated with filtrate, please discard the filtrate and put the nucleic acid purification column back into the 2 ml centrifuge tube and empty it at the highest speed for 1 minute before taking it out. Nucleic acid purification columns perform this step.

* If the centrifuge does not have a leak-proof lid, please change the centrifugation conditions to 8000 rpm for 1 minute to prevent the tube lid from falling off and damaging the centrifuge.

8.       Discard the purification column, and the eluted DNA can be used immediately for various molecular biology experiments; or the DNA can be stored at -20°C for later use.