BIOG stool DNA extraction kit instructions

Transportation conditions: normal temperature transportation

Item No.: 51031: 50 reactions

51032: 100 reactions

Storage conditions: store at room temperature (15-25 ° C) digestive solution at -20 ° C, lysate I should be stored at 4 ° C

Features

Kit composition

product description

The BIOG DNA Stool Kit is designed by our research and development team to extract genomic DNA from feces and is suitable for solid or liquid stool samples from different sources. The BIOG DNA Stool Kit uses the latest ion-exchange membrane technology and has repeatedly optimized the lysate and eluent formulations. The extracted DNA is of higher purity and minimizes the contamination of proteins, pigments, lipids and impurities. PCR, real-time PCR, various enzyme digestion tests, and the like.

Steps:

Please prepare yourself: absolute ethanol, PBS, 1.5mL centrifuge tube.
Take out the supernatant and washing solution as follows:
a) Precipitate: 4.5 mL was added to 25.5 mL of absolute ethanol; 9 mL was added to 51 mL of absolute ethanol.

b) Washing solution: 9 mL of 21 mL of absolute ethanol was added; 18 mL of 42 mL of absolute ethanol was added.

c) If the prepared precipitate is precipitated, it can be dissolved at 37 ° C, shaken and used.

Take 200mg of feces for the test tube (if the stool sample is liquid, take 200μL in the test tube), add 2mL PBS and mix well by shaking, centrifuge at 300g for 5 minutes, collect the supernatant; take the collected supernatant 1mL in 1.5mL centrifuge tube The cells were centrifuged at 12,000 rpm for 5 minutes, and the supernatant was discarded, resuspended in 1 mL of PBS, shaken and mixed, and centrifuged at 12,000 rpm for 5 minutes to collect a precipitate.

200 μL of Lysate I was added, the pellet was suspended, and left at 37 ° C for 30 minutes.

Add 200 μL of Lysis Buffer II, 20 μL of the digestive juice, mix well and shake well, and boil at 56 ° C for 10 minutes.

Add 600 μL of the precipitate and mix gently by inversion. If there is a semi-transparent suspension, it will not affect the DNA extraction and subsequent experiments.

The adsorption column was placed in a collection tube, and 600 μL of the above solution was transferred to the adsorption column, allowed to stand for 2 minutes, centrifuged at 12,000 rpm and 4 ° C for 1 minute, and the waste liquid in the collection tube was discarded.

The remaining 400 μL of the liquid was transferred into the above adsorption column, and operation step 7 was repeated.

Put the adsorption column back into the collection tube, add 500 μL of the washing solution to the adsorption column, centrifuge at 12,000 rpm for 4 minutes at 4 ° C, and discard the waste liquid in the collection tube.

Place the column back into the collection tube and centrifuge at 12,000 rpm for 4 minutes at 4 ° C to remove any remaining wash solution.

Remove the adsorption column, place it in a new 1.5 mL centrifuge tube, add 30-100 μL of the eluate, let stand for 3 minutes, centrifuge at 12,000 rpm for 4 minutes at 4 ° C, and collect the DNA solution. The extracted DNA can be used in the next experiment or at -20 °C.

Precautions:

1. The liquid and washing liquid contain irritating chemicals. Please take protective measures during operation to avoid direct contact with the skin and prevent inhalation of nose and mouth. If you accidentally get into skin or eyes, rinse immediately with water or saline. Seek medical attention if necessary.

2. If the lysate is precipitated as a white floc, it is a normal phenomenon and can be dissolved in a 37 ° C water bath.

Storage, transportation and expiration date

This kit can be transported at room temperature, stored at room temperature (15-25 °C), valid for 12 months. Store digestive solution at -20 °C to avoid repeated freezing and thawing. Store lysate I at 4 °C.

QC

This product has passed strict quality inspection and proved that there is no biological pollution.

Note This product is for scientific research only.