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Pretreatment solution for PCR detection sample

A technology for pretreatment of liquid and samples, applied in the field of molecular biology, can solve problems such as complex operation, and achieve the effects of convenient operation, lower detection costs, and lower transportation costs.

Active Publication Date: 2021-03-26
ZYBIO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention provides a pretreatment solution for PCR detection samples to solve the complex operation of the prior art, realize the purpose of simultaneously preserving the sample and extracting nucleic acid in the pretreatment solution, and simplify the operation

Method used

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  • Pretreatment solution for PCR detection sample
  • Pretreatment solution for PCR detection sample
  • Pretreatment solution for PCR detection sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Study on Stability of Pretreatment Solution 1

[0053]The influence of temperature and time: Collect hepatitis B virus with sampling test, add to pretreatment solution 1, store at 25°C, 4°C, -20°C, -70°C for 12 hours, 24 hours, 48 ​​hours, 72 hours , followed by viral nucleic acid extraction and amplification.

[0054] The results are shown in Table 2

[0055] The preservation effect is shown in Table 2, and the results show that the Ct value gradually increases with the increase of time, indicating that the integrity of the virus decreases with the increase of time. In the detection time range, the temperature has little effect on the effect of the pretreatment solution, indicating that the pretreatment solution is not affected by temperature and can be stored stably within the range of -70°C to 25°C.

[0056] Table 2 The influence of temperature and time on pretreatment solution 1

[0057]

Embodiment 2

[0059] Collect hepatitis B virus with sampling test, add to pretreatment solution 2, store at 25°C, 4°C, -20°C, -70°C for 12 hours, 24 hours, 48 ​​hours, 72 hours, and then carry out virus nucleic acid extraction and amplification.

[0060] The results are shown in Table 3. The results show that the Ct value gradually increases with time, indicating that the integrity of the virus decreases with time.

[0061] However, within the detection time range, the temperature has little influence on the effect of the pretreatment solution. As the temperature rises, the difference in Ct value at the same time is not very large, indicating that the pretreatment solution is not affected by temperature and can be used in Stable storage within the range of -70℃~25℃.

[0062] The influence of table 3 temperature and time on pretreatment liquid 2

[0063]

Embodiment 3

[0065] Collect hepatitis B virus with sampling test, add to pretreatment solution 3, store at 25°C, 4°C, -20°C, -70°C for 12 hours, 24 hours, 48 ​​hours, 72 hours, and then carry out virus nucleic acid extraction and amplification.

[0066] The results are shown in Table 4, and the results show that the Ct value gradually increases with the increase of time, indicating that the integrity of the virus decreases with the increase of time.

[0067] However, within the detection time range, the temperature has little influence on the effect of the pretreatment solution. As the temperature rises, the difference in Ct value at the same time is not very large, indicating that the pretreatment solution is not affected by temperature and can be used in Stable storage within the range of -70℃~25℃.

[0068] The influence of table 4 temperature and time on pretreatment liquid 3

[0069]

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Abstract

The invention relates to the technical field of molecular biology, and discloses a pretreatment solution for a PCR detection sample. According to the pretreatment solution, an anionic polymer is added, sample preservation under the normal temperature condition and nucleic acid extraction under the heating condition are achieved through the pretreatment solution, magnetic bead addition, protease Kaddition or physical crushing, centrifuging and the like do not need to be adopted for splitting samples, operation is easy, the extraction effect is good, and the pretreatment solution has importantsignificance on development and application of nucleic acid extraction.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a pretreatment liquid for PCR detection samples. Background technique [0002] Magnetic bead nucleic acid detection is widely used in medical diagnosis because of its high sensitivity. The detection process mainly includes magnetic bead nucleic acid extraction and PCR detection. Before magnetic bead nucleic acid extraction, the sample needs to be processed first, and the sample is temporarily stored in the preservation solution; the usual magnetic bead nucleic acid extraction can be divided into four steps: (1) lysis; (2) binding; (3) washing; (4) Separation. Specifically, cells or tissues release DNA / RNA under the action of lysate, and the surface-modified superparamagnetic silica nano-magnetic beads specifically bind to the released DNA / RNA to form a nucleic acid-magnetic bead complex. Subsequently, washing liquid is added to the nucleic acid-magnetic bead complex ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12Q1/70C12Q1/686C12R1/93
CPCC12Q1/6806C12Q1/706C12Q1/701C12Q1/686C12Q2527/125C12Q2531/113C12Q2521/107C12Q2521/531
Inventor 董璐陈威张天张越
Owner ZYBIO INC
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