A method for detecting the length of telomere DNA based on electrotransfer and vacuum transfer
A transfer method, telomere technology, applied in the field of DNA detection, can solve the problems of low transfer efficiency, poor repeatability, and long time consumption, and achieve the effect of accurate results and fast transfer speed
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Embodiment 1
[0046] Such as Figure 1-2 As shown, a telomere DNA transfer device includes a negative pressure box 1 , a positive plate 2 , a DNA transfer assembly 3 and a negative plate 4 .
[0047] The negative pressure box 1 is a hollow box body, a mounting groove 101 is provided at the top opening, and an air suction port 102 is provided at the side.
[0048] The positive plate 2 is placed in the installation groove 101, the DNA transfer assembly 3 and the negative plate 4 are placed on the positive plate 2 in sequence, and the layers are closely attached to each other.
[0049] The positive plate 2 and the negative plate 4 are respectively connected to the positive and negative poles of an external power supply. The preparation method of the positive plate 2 and the negative plate 4 is as follows: mix graphene powder and 100-mesh silica sand powder at a ratio of 50:1, put them in a mold and sinter at high temperature to form an electrode plate with air holes with a diameter of 30-50 μ...
Embodiment 2
[0054] As shown in 2-5, a telomere DNA transfer device includes a negative pressure box 1 , a support plate 5 , a positive plate 2 , a DNA transfer assembly 3 and a negative plate 4 .
[0055] The negative pressure box 1 is a hollow box body, a mounting groove 101 is provided at the top opening, and an air suction port 102 is provided at the side.
[0056] The support plate 5 is a stainless steel mesh plate, on which a plurality of air vents 501 are evenly distributed; the support plate 5 is placed in the installation groove 101 .
[0057] The positive plate 2, the DNA transfer assembly 3 and the negative plate 4 are sequentially placed on the support plate 5, and the layers are closely attached to each other.
[0058] The positive plate 2 and the negative plate 4 are respectively connected to the positive and negative poles of an external power supply. The preparation method of the positive plate 2 and the negative plate 4 is as follows: mix graphene powder and 100-mesh sili...
Embodiment 3
[0063] The detection method of embodiment 3 telomeric DNA length
[0064] 1. Preparation of telomeres (genomic DNA was extracted using Qiagen69506 kit)
[0065] (1) Culture cells in a 10cm cell culture dish, discard the cell culture medium, wash twice with PBS, digest with trypsin at 37°C for 3 minutes, collect in a 15mL centrifuge tube, centrifuge at 200g for 3 minutes, discard the supernatant, wash the cells twice with PBS, Transfer to a 1.5mL centrifuge tube.
[0066] (2) Add 200 μL of BufferTL, mix by pipetting, then add 20 μL of proteinase K, shake and digest in a 57°C water bath for 30-60 minutes until the cells are completely lysed.
[0067] (3) Add 220 μL BufferBL, vortex to mix, shake and stand in a 50°C water bath for 10 minutes.
[0068] (4) Add 220 μL absolute ethanol, and vortex to mix.
[0069] (5) Put the DNA adsorption column in the kit into a 2mL collection tube, add all the liquid obtained in the previous step to the adsorption column, and centrifuge at 13...
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