Blood DNA Kit Urine Genome DNA extraction Kit (column type)

User Manual

Product Name

Common name: Blood DNA Kit Urine Genome DNA extraction Kit (column type)

Trade name: Blood DNA Kit Urine Genome DNA extraction Kit (column type)
Model: ZT-003

Intended use

This kit uses a centrifugal adsorption column that can specifically bind nucleic acids and a unique buffer system to extract genomic DNA from urine samples. The silicon matrix material used in the centrifugal adsorption column can efficiently and specifically adsorb nucleic acids, and combined with a unique buffer system, it can maximally remove impurities such as proteins and other organic compounds in cells. The extracted nucleic acid fragments are complete, highly purified, and of stable and reliable quality. The nucleic acid purified using this kit is suitable for various conventional experiments such as enzyme digestion, PCR, library construction, Southern hybridization, and experiments that require high-quality nucleic acid such as chip hybridization and high-throughput sequencing. This product is expected to be used by professionals, such as technicians and doctors trained in molecular biology technology.
  

Inspection principle

1. High quality: Unique lysis buffer system, purified DNA with high concentration, high purity, and good integrity, meeting experimental requirements such as chip hybridization and high-throughput sequencing.

2. Fast and non-toxic: Adopting the principle of silica gel membrane adsorption, without the need for phenol chloroform.

Precautions (please be sure to read this note before using this reagent kit):

1. The sample should avoid repeated freeze-thaw, otherwise it may result in smaller extracted DNA fragments and a decrease in extraction volume.

If there are precipitates in cracking solution 1 and 3, they can be dissolved again in a 70 ℃ water bath, shaken well, and then used.

3. Balance the sample and eluent 3 to room temperature.

4. All centrifugation steps are performed at room temperature.
  

【 Main components 】

Items that customers need to prepare by themselves include: 96~100% ethanol, 1 M DTT (Dithiothreitol); After packaging, store at -20 ℃, thaw before use, PBS, gun head with filter element, metal bath that can be heated to 58 ℃, centrifuge (1.5ml and 2ml centrifuge tubes can be placed), vortex oscillator.
  

Storage conditions and validity period

The reagent kit can be stored for 12 months under dry conditions at room temperature (15-25 ℃). If the room temperature exceeds 25 ℃, adsorption column 1 and protease can be stored at 2-8 ℃.
  

Reagent preparation

Before use, add 96-100% ethanol to Lotion 3 and Lotion 4, and add 17ml 96-100% ethanol to Lotion 3 bottles; Add 21ml of 96-100% ethanol to 4 bottles of washing solution. Add 1.5ml of protease solution to the protease bottle for dissolution (store the dissolved protease at 2-8 ℃).
  

[Operation Steps]

Centrifuge 6000g of 1.1-10ml urine sample for 2 minutes.

2. Remove the supernatant and add 500 μ L eluent 3, vortex oscillation for 5 seconds.

Centrifuge 3.6000g for 2 minutes, remove the supernatant, and add 300 μ Cracking liquid 1 and 25 μ L protease, vortex oscillation for 10 seconds.

Note: Because urine may contain sperm cells, DTT can lyse sperm cells, so add 20 μ 1 M DTT of l can improve sensitivity.

4. Place it in a whirlpool shaker with a temperature bath of 58 ℃ and shake at 900 rpm for 1 hour. If using a dry or water bath that cannot be shaken well, mix well every 15 minutes.

5. Instantaneous centrifugation, adding 300 μ Cracking liquid 3 and 50 μ Ethanol (96~100%), vortex oscillation for 10 seconds.

Note: If the room temperature exceeds 25 ° C, cool the ethanol on ice before adding it to the tube.

6. Instantaneous centrifugation, add to the adsorption column, centrifuge at a speed of over 6000g for 1 minute, place the adsorption column in a clean 2 ml collection tube, and discard the collection tube containing the effluent.

Note: 6000g centrifugation will only reduce noise, and centrifugation at a higher speed will not affect DNA extraction.

7. Add 500 μ Centrifuge at a rate of 6000g of washing solution 3 (with ethanol added) for 1 minute, place the adsorption column in a clean 2 ml collection tube, and discard the collection tube containing the effluent.

8. Add 500 μ Centrifuge solution 4 (with ethanol added) at a rate of 6000g for 1 minute, place the adsorption column in a clean 2 ml collection tube, and discard the collection tube containing the effluent.

Centrifuge at a speed of 9.20000 g for 3 minutes.

10. Replace the centrifuge tube and add 20 drops to the middle position of the adsorption membrane μ L-50 μ Elute 3, warm bath at room temperature for 2 minutes, centrifuge at a rate of 20000gMinutes.