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Excrement sample DNA gene mutation kit

A fecal sample and kit technology, applied in recombinant DNA technology, DNA/RNA fragment, microorganism determination/inspection, etc., can solve the problem of poor detection effect, low actual specificity and accuracy, and inability to control the reaction process well and other problems, to achieve the effect of good detection effect, actual specificity and high accuracy

Pending Publication Date: 2022-01-11
苏州丽纳芯生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, when the stool sample DNA gene mutation kit is actually used, due to the lack of good primer probe design, the actual detection method is single, and the overall actual detection effect is not good. Control the reaction process to obtain accurate and reliable experimental results, but the actual specificity and accuracy are low

Method used

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  • Excrement sample DNA gene mutation kit
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  • Excrement sample DNA gene mutation kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0034] The embodiment of the present invention provides a stool sample DNA gene mutation kit, comprising the following specific steps:

[0035] S1. For KRAS mutation detection and β-actin internal reference, nucleic acid extraction is first required. Nucleic acid extraction kit is used to extract nucleic acid from feces in preservation solution. Nanodrop is used to detect A26 / A280, and the nucleic acid with Fe3O4 magnetic microspheres as the carrier is used The extraction method shows its unique convenience and quickness. The magnetic microspheres do not need centrifugation during the nucleic acid purification process, avoiding cross-contamination, and the extraction results are reproducible and easy to operate;

[0036] S2. Detected by the peptide nucleic acid clamp (PNA) method, 98% of the mutations of the KRAS gene occurred at codons 12 and 13 of exon 2, including G12C, G12S, G12R, G12V, G12D, G12A and G13D;

[0037] S3. Use PNA technology for detection, and KRAS primers ar...

Embodiment 2

[0047] The difference between this example and Example 1 is that: PNA technology is used for detection, and the KRAS primers are as follows:

[0048] .

[0049] Preferably, different mutation type probes are as follows:

[0050] .

[0051] Preferably, fluorescent signal:

[0052] 5' 3' FAM BHQ-1 JOE TAMRA CY3 BHQ-2

Embodiment 3

[0054] 1. Kras mutation

[0055] (1) Enrichment of known low-abundance DNA mutation detection technology, peptide nucleic acid clamp PCR (PNA-mediated PCR): PCR technology based on peptide nucleic acid (peptide nucleic acid, PNA), was first established and used by Egholm et al. Peptide nucleic acid (PNA) is a DNA analog with a polypeptide-like backbone. The main chain backbone is composed of N(2-aminoethyl)-glycine and nucleic acid bases connected by methylene carbonyl51. PNA can specifically bind to DNA Or RNA hybridization to form a stable complex. The reason why PNA can specifically clamp the amplification of specific alleles: firstly, PNA cannot be used as a primer; secondly, PNA cannot be degraded by nuclease and protease. Moreover, the thermostability of PNA-DNA and PNA-RNA is higher than that of corresponding DNA.

[0056] (2) The extraction method is divided into centrifugation method and magnetic bead method. Choose a better method based on the nanodrop results. T...

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Abstract

The invention provides an excrement sample DNA gene mutation kit, and relates to the technical field of kits. The excrement sample DNA gene mutation kit comprises KRAS mutation detection and beta-act i n internal reference, firstly, nucleic acid extraction work needs to be performed, the nucleic acid extraction kit is adopted, excrement in a preservation solution is subjected to nucleic acid extraction and Nanodrop detection of A26 / A280, and a nucleic acid extraction method with Fe3O4 magnetic microspheres as a carrier is adopted. By adopting good primer probe design, adopting a PNA technology for detection and arranging different mutation type probes, actual detection modes are diversified, and the overall actual detection effect is good; and in addition, through scientific experimental steps, experimental modes and experimental conditions, the reaction process can be well mastered, then accurate and reliable experimental results are obtained, and the actual specificity and the accuracy are high.

Description

technical field [0001] The invention relates to the technical field of kits, in particular to a stool sample DNA gene mutation kit. Background technique [0002] The kit is a box used to contain chemical reagents such as chemical components, drug residues, and virus types. In the detection of methylation and gene mutation in colorectal cancer, stool sample DNA gene mutation kit is commonly used. Stool gene detection is convenient for sampling and can minimize the probability of discomfort during the examination process. [0003] At present, when the stool sample DNA gene mutation kit is actually used, due to the lack of good primer probe design, the actual detection method is single, and the overall actual detection effect is not good. Control the reaction process to obtain accurate and reliable experimental results, but the actual specificity and accuracy are low. [0004] To this end, we have developed a new DNA gene mutation kit for stool samples. Contents of the inve...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6886C12Q1/6858C12Q2600/156C12Q2525/107C12Q2531/113C12Q2563/107C12Q2523/308
Inventor 张琬祺谭生伟何木兰
Owner 苏州丽纳芯生物科技有限公司
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