Micro ribonucleic acid (RNA) molecular mark for diagnosing gestational diabetes and detection kit thereof
A technology of gestational diabetes and kits, applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial determination/inspection, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0015] Example 1. Microarray analysis of miRNA expression profile changes in the placenta of pregnant women with gestational diabetes.
[0016] Take 20 placentas from patients with gestational diabetes and 20 placentas from normal pregnant women as controls. Tissue RNA was extracted as follows.
[0017] ①Take fresh placenta samples, mix 20 samples, freeze in liquid nitrogen, grind with a mortar until the tissue becomes powdery (use liquid nitrogen to keep the sample at a low temperature during the grinding process), add an appropriate amount of sample powder to 1ml of TRIZOL, and store at room temperature Leave it for 1h.
[0018] ②Add 200 μl of chloroform to TRIZOL, shake vigorously, let stand for 2 minutes, centrifuge at 4°C, 12000 rpm for 15 minutes.
[0019] ③ Remove the supernatant, add an equal volume of isopropanol to the supernatant, shake vigorously, let stand at room temperature for 10 minutes, centrifuge at 12000 rpm at 4°C for 10 minutes.
[0020] ④ Discard the ...
Embodiment 2
[0025] Embodiment 2 uses real-time PCR technology to verify the result of the chip
[0026] Take 18 placentas from patients with gestational diabetes and 18 placentas from normal pregnant women as controls. Tissue RNA was extracted as follows.
[0027] ① Take a fresh placenta sample, freeze it in liquid nitrogen, grind it with a mortar until the tissue becomes powdery (use liquid nitrogen to keep the sample at a low temperature during the grinding process), add an appropriate amount of sample powder to 1 ml of TRIZOL, and place it at room temperature for 1 hour.
[0028] ②Add 200 μl of chloroform to TRIZOL, shake vigorously, let stand for 2 minutes, centrifuge at 4°C, 12000 rpm for 15 minutes.
[0029] ③ Remove the supernatant, add an equal volume of isopropanol to the supernatant, shake vigorously, let stand at room temperature for 10 minutes, centrifuge at 12000 rpm at 4°C for 10 minutes.
[0030] ④ Discard the supernatant, and place the precipitate in a ventilated place t...
Embodiment 3
[0040] Example 3 Using real-time PCR technology to detect the expression level of miR-508 in the serum of pregnant women
[0041] The serum of 18 cases of gestational diabetes patients and 18 cases of normal pregnant women were taken as controls. RNA was extracted as follows.
[0042] ①Take 500ul of fresh serum sample, add 500ul of TRIZOL to the serum, shake vigorously for 30s, and place at room temperature for 5min.
[0043] ②Add 200ul of isopropanol to TRIZOL, invert and mix well, shake vigorously for 2 minutes until the liquid is transparent, and let stand at room temperature for 5 minutes.
[0044] ③ 4°C, 12000rpm, centrifuge for 10min, and transfer the supernatant to a new eppendorf tube.
[0045] ④Add 500ul chloroform to the supernatant, mix by inverting, shake vigorously for 1min, and let stand at room temperature for 5min.
[0046] ⑤ 4°C, 12000rpm, centrifuge for 10min, and transfer the supernatant to a new eppendorf tube.
[0047] ⑥ Add 0.75 times the volume of is...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More 