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A kind of cervical cell lysing kit and lysing method

A cervical cell and lysis buffer technology, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve problems such as inability to amplify experiments, inability to fully dissociate DNA, and substandard DNA quality and concentration, and achieve a good amplification curve Effect

Active Publication Date: 2020-07-17
DALIAN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the large number of interference items in the sample, the existing commercial kits cannot fully dissociate the DNA from the cervical TCT preservation solution. The quality and concentration of the extracted DNA are not up to standard, and subsequent amplification experiments cannot be performed. Unable to obtain accurate genetic test results

Method used

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  • A kind of cervical cell lysing kit and lysing method
  • A kind of cervical cell lysing kit and lysing method
  • A kind of cervical cell lysing kit and lysing method

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Cervical Epithelial Cell Lysis Kit

[0038] Preparation of lysis buffer:

[0039]

[0040]

[0041] Proteinase K: 20mg / ml.

[0042] Cell Lysis Method:

[0043] In the process of clinical cervical TCT detection, add 10ml of the preservation solution containing cervical cells into an equal volume of PBS to wash, centrifuge at 4600g for 60min, carefully remove the supernatant, and collect the precipitate;

[0044] Add 1ml of the above lysis buffer, pipette to mix the precipitate, add 50μl proteinase K, and digest at 55°C for 12h;

[0045] Vortex for 50 seconds, and continue to digest at 55°C for 2 hours until the cells are completely lysed.

[0046] After cell lysis, DNA extraction was performed according to step 10 in the paraffin-embedded tissue DNA extraction kit (Beijing Tiangen / DP331-02) from step 10 to completion (kit instruction manual), specifically including the following steps: aliquot the lysed liquid Add 220μl GB to 1.5ml EP tube, 220μl per tube, vort...

Embodiment 2

[0064] Cervical Epithelial Cell Lysis Kit

[0065] The lysis buffer uses water as the solvent and includes the following components: 200mM Tris-HCl, 50mM EDTA, 0.1wt% SDS and 200mM NaCl

[0066] Proteinase K: 20mg / ml.

[0067] Cell Lysis Method:

[0068] In the process of clinical cervical TCT detection, add 12ml of the preservation solution containing cervical cells to 2 times the volume of PBS to wash, centrifuge at 4600g for 60min, carefully remove the supernatant, and collect the precipitate;

[0069] Add 1ml of the above lysis buffer, pipette to mix the precipitate, add 50μl proteinase K, and digest at 55°C for 12h;

[0070] Vortex for 50 seconds, and continue to digest at 55°C for 2 hours until the cells are completely lysed.

[0071]After the cells were lysed, the DNA extraction was performed according to step 10 in the paraffin-embedded tissue DNA extraction kit (Beijing Tiangen / DP331-02) from step 10 to completion (the kit instructions are detailed), specifically i...

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Abstract

The invention provides a cervical cell lysis kit and a cell lysis method, belonging to the technical field of cell lysis. The kit includes a lysis buffer and proteinase K. The lysis buffer comprises the following components in concentration: 10-500mM of Tris-HCl, 1-80 mM of EDTA, 0.05-2 wt% of SDS and 100-250 mM of NaCl. The cell lysis method comprises the following steps: after cleaning a cervical cell sample solution with PBS buffer, uniformly mixing the solution with lysis buffer, adding proteinase K, and digesting at 53-58 DEG C for 10-14 hours; After 30-60 s of vortex vibration, digestingthe cells at 53-58 DEG C for 1.5-2.5 h to complete cell lysis. The kit and the lysis method can fully lyse cervical cells in the sample, dissociate high-concentration and high-quality DNA, and facilitate subsequent DNA extraction and amplification.

Description

technical field [0001] The invention belongs to the technical field of cell lysis, in particular to a cervical cell lysis kit and a lysis method. Background technique [0002] Cervical cancer is one of the common malignant tumors that threaten women's health. Effective screening and early diagnosis can significantly reduce the incidence and mortality of cervical cancer. Cervical liquid-based thin-layer cytology testing (TCT) has become one of the common methods for cervical cancer screening and cytological diagnosis. TCT detection is a technology that preserves the exfoliated cells in the preservation solution, and uniformly disperses and adheres the cells to the glass slide through a special film-making instrument for staining and observation. When clinically collecting samples, a special cervical brush should be inserted into the cervix to collect exfoliated epithelial cells from the cervix and cervical canal, and the brush head of the cervical brush should be placed in a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 肖晶李楠
Owner DALIAN MEDICAL UNIVERSITY
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