Abbexa cell free DNA kit instructions

AI Meijie Abbexa No cell  DNA  The kit is used to purify acellular cells by enzymatic hydrolysis of samples and specific adsorption of silica magnetic beads  DNA  The ideal solution . Apply to from  0.5-10 ml  Separation and purification of high-quality free from serum or plasma  DNA. Extracted  DNA  Can be used for  PCR、qPCR  and  NGS, It can also be used in nucleic acid extraction system based on magnetic beads .

AI Meijie Abbexa Kit components ：

Binding buffer ：120  ml

Cleaning buffer ：30  ml

Wash buffer ：24  ml

Elution buffer ：4 ml

protease  K (20 mg/ml)：3 × 1 ml

20% SDS：6  ml

Magnetic acellular beads ：2  ml

AI Meijie Abbexa  No cell  DNA Basic parameters of the kit ：

The goal is   free  DNA

Keep in storage   Place the magnetic beads on  2-8°C  Store for up to a year , Don't freeze . Store all other reagents at room temperature .

Instructions   Reagent preparation ：

Work with buffer ： stay  120  Add... To ml of binding buffer  40  Ml isopropyl alcohol , Preparation work combined with buffer solution .

Clean the working buffer ： stay  30  Add... To ml of cleaning buffer  30  Ml isopropyl alcohol , Prepare cleaning buffer solution .

Working wash buffer ： take  96  Ml isopropyl alcohol is added to  24  Ml of washing buffer to prepare working washing buffer .

notes ：

Use the front vortex beads .

Use sterile 、 Nucleic acid free and nuclease free centrifuge tubes and pipettes .

free  DNA  The component content of J low , It is recommended to use a low nucleic acid adsorption centrifuge tube for plasma storage 、DNA  Extract and  DNA  Store .

Avoid reagents 、 Sample and separated  DNA  Repeated freeze-thaw cycles .

According to the following table, in  15 ml  or  50 ml  The reaction system is prepared in a centrifuge tube .

Spare parts   Plasma volume

0.5  ml  1  ml  2  ml  4 ml  10  ml

20%  Safety data sheets  25 Microlitre  50 Microlitre  100  Microlitre  200  Microlitre  500  Microlitre

protease K 15 Microlitre  30 Microlitre  60 Microlitre  120  Microlitre  300  Microlitre

Work binding buffer  0.75  ml  1.5  ml  3  ml  6 ml  15  ml

Magnetic acellular beads  10 Microlitre  20 Microlitre  40 Microlitre  80 Microlitre  200  Microlitre

Testing procedure ：

1、 Establish the reaction system according to the above table .

2、 Vortex tube  15  second , Then place at room temperature  20  minute . During this time, turn the test tube upside down  3-5  Time .

3、 Magnetic separation ： Place the centrifuge tube on the magnetic frame , Then gently rotate the centrifuge tube left and right by hand . When the magnetic beads begin to gather towards the tube wall , Reverse the spin direction , repeat  2-3  Time . Make sure that any beads on the cover collect on the pipe wall . Standing  2  Minutes and make sure that all beads gather on the tube wall .

4、 Discard the supernatant from the other side of the bead , Be careful not to remove the magnetic beads themselves . Remove the test tube from the magnetic holder and add  1 ml  Working cleaning buffer （ Contains isopropyl alcohol ）. Vortex  15  second , Then move to the new  1.5 ml  In the centrifuge tube . If the magnetic beads remain in the original tube , Transfer the supernatant back to the original tube , wash , And then move on to  1.5 ml  In the centrifuge tube . Repeat step  3  Perform another magnetic separation .

5、 Discard the supernatant . Remove the test tube from the magnetic holder , Join in  1 ml  Working wash buffer （ Contains isopropyl alcohol ）. Vortex  15  second , Then repeat the steps  3  Perform another magnetic separation .

6、 Repeat step  5.

7、 Discard the supernatant , Including any liquid on the lid . It is recommended to use a smaller pipette suction head to completely remove the supernatant .

8、 Let stand and air dry  5-10  minute .

9、 Add elution buffer according to the following table

Spare parts   Original plasma volume

0.5  ml  1  ml  2  ml  4 ml  10  ml

Elution buffer  20 Microlitre  30 Microlitre  50 Microlitre  75 Microlitre  200  Microlitre

Vortex tube  5  minute .

10、 Place the centrifuge tube on the magnetic support . Transfer the liquid to a new  1.5 ml  Low nucleic acid adsorption in centrifuge tube , Be careful not to remove the magnetic beads themselves .

11、 To separate  DNA  Stored in the -20°C.