A method for rapid extraction of rna using an animal and plant tissue ablation instrument based on the principle of ultrasound

A tissue ablation, animal and plant technology, applied in the field of rapid RNA extraction, can solve the problems of incomplete fragmentation, difficult to inhibit endogenous RNase, complicated operation, etc., to avoid incomplete tissue fragmentation, shorten the extraction time, and improve the extraction efficiency. Effect

Active Publication Date: 2020-10-09
CHENGDU DAOSHENG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The liquid nitrogen grinding method requires a large tissue block, and it is difficult to completely break up the tissue block; the homogenization method is difficult to completely inhibit the activity of endogenous RNase, and it is easy to cause endogenous degradation; the disadvantage of the repeated freeze-thaw method is that it is difficult to Thoroughly lyses large tissues, and endogenous RNase activity is difficult to inhibit
Therefore, the RNA extraction of animal-derived tissue blocks has difficulties such as complicated operation, incomplete fragmentation, and difficulty in inhibiting the activity of endogenous RNA, which makes it difficult to guarantee the quality and yield of RNA extraction.

Method used

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  • A method for rapid extraction of rna using an animal and plant tissue ablation instrument based on the principle of ultrasound
  • A method for rapid extraction of rna using an animal and plant tissue ablation instrument based on the principle of ultrasound
  • A method for rapid extraction of rna using an animal and plant tissue ablation instrument based on the principle of ultrasound

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] like Figure 1-6 As shown, the animal and plant tissue ablation instrument based on the principle of ultrasound in this embodiment includes a power supply, a DCDC conversion unit, a main control unit, a DCDC power adjustment unit, a drive unit, a transformer unit, a resonance unit, an ultrasonic transducer and a sampling unit .

[0098] The power supply converts the voltage through the DCDC conversion unit and the DCDC power adjustment unit to the animal and plant tissue ablation instrument; the main control unit outputs the first PWM signal to control the DCDC power adjustment unit to output adjustable voltage, and collects the output terminal of the transformation unit Feedback from the sampling unit of the loop voltage and current, output two second PWM signals with complementary duty ratios to the drive unit and transform the voltage through the transformer unit, and then adjust the resonance through the resonance unit set in the output circuit of the transformer un...

Embodiment 2

[0131] On the basis of Embodiment 1, this embodiment further discloses an animal and plant tissue ablation instrument based on the principle of ultrasound. In a specific application, the main control unit of the present invention also includes an auxiliary chip U1 and auxiliary chip peripheral circuits, and the auxiliary chip U1 The model number is STM32F031G4U6.

[0132] The features of STM32F031G4U6 are: Core: 32 bit -M0 CPU with frequency up to 48MHz; Storage: 16 to 32KB of flash memory, 4KB of SRAM with hardware parity; CRC calculation unit; reset and power management Digital and I / O power supply: 2.0 to 3.6V, analog power supply: VDDA = from VDD to 3.6V, power-on / power-down reset (POR / PDR), programmable voltage detector (PVD), low-power modes: sleep, stop and support, VBAT power supply for RTC and backup registers; Clock Management: 4 to 32MHz crystal oscillator, 32kHz oscillator for RTC with calibration, internal 8MHz RC with x6PLL option, internal 40kHz RC oscillator...

Embodiment 3

[0147] This embodiment provides an extraction solution, including 1.6 mol / L guanidine thiocyanate and 1 mol / L guanidine hydrochloride, and the pH of the extraction solution is adjusted to 4 by acetic acid. The extract is applied to the preserved tissue or cells.

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Abstract

A method for rapidly extracting RNA using an ultrasonic principle-based animal and plant tissue ablation instrument, comprising: 1) first preserving tissues or cells using an extracting liquid A and then adding an extracting liquid B into the tissues or cells preserved using the extracting liquid A, and uniformly mixing to obtain a mixture; 2) performing ultrasonic crushing on the mixture when extracting RNA from the tissues; 3) directly performing the operation in step 5) below when extracting RNA from the cells; 4) uniformly mixing the mixture subjected to ultrasonic crushing by inversion; 5) in the mixture of RNA extracted from the cells in step 3) or the mixture of step 4), adding chloroform for centrifugation, and taking an upper layer of liquid; 6) adding isopropanol into the upper layer of liquid, and then performing centrifugation to obtain a precipitate; 7) adding an ethanol solution into the precipitate, uniformly mixing, centrifuging, and discarding the liquid; 8) finally, adding water for dissolution.

Description

technical field [0001] The invention belongs to the technical field of biological preparations, and in particular relates to a method for rapidly extracting RNA by using an animal and plant tissue ablation instrument based on the principle of ultrasound. Background technique [0002] In modern molecular biology experiments and clinical molecular diagnosis, it is often necessary to isolate and purify RNA from tissues and cells, and the quality of RNA often affects the success or failure of molecular biology experiments such as cDNA library, RT-PCR and Northern Blot. [0003] The commonly used method is to first protect the tissue with a tissue protection solution, and then use the extraction solution to extract the RNA in the tissue or cells. Commonly used protection methods include liquid nitrogen protection and protection solution protection, and the commonly used protection solutions include RNA later reagents and RNA safety reagents. [0004] Among them, the liquid nitro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003C12Q2523/301
Inventor 赵仲玖廖东升邱坤赵伟
Owner CHENGDU DAOSHENG BIOTECH CO LTD
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