Solid phase polymerase chain reaction (PCR) kit for chemotherapeutic medication guide gene detection
A technology for medication guidance and gene detection, applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., can solve the problem of high infection risk, achieve large detection throughput, avoid non-specific amplification, and improve detection sensitivity and specificity. sexual effect
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Embodiment 1
[0023] Composition of solid-phase PCR detection kit for guidance of 10 kinds of chemotherapeutic drugs
[0024]The kit consists of a PCR reaction mixture and 12 strip tubes or a 96-well PCR reaction plate, in which the PCR reaction mixture is 5×PCR Buffer (Tris-HCl pH8.5 100mM, KCl 500nM, MgCl2 15nM), dNTPs ( 10mMeach), Mg2+, 12 pairs of primers and hot start DNA polymerase.
[0025] The kit is to seal a layer of paraffin wax mixture on the surface of the above-mentioned PCR reaction mixture, and the paraffin wax mixture is formed by mixing solid paraffin and liquid paraffin according to 1:8 (W / V).
[0026] The reaction system (25ul) of the kit is: PCR reaction solution 23ul, template 2ul.
Embodiment 2
[0028] Kit operation and result determination
[0029] 1. Genomic DNA extraction
[0030] Using paraffin-embedded tissue DNA extraction kit (Tiangen Biochemical Technology, product number: DP331) to extract cancer tissue DNA according to the kit instructions.
[0031] 2. Preparation of reaction system
[0032] Take Chemotherapy Medication Guidance Solid-Phase PCR Reaction Kit 12-tube strip or 96-well PCR reaction plate, and add 2ul of extracted DNA.
[0033] 3. PCR amplification
[0034] Put the PCR tube into an ordinary PCR machine and perform PCR amplification according to the instructions recommended.
[0035] 4. Analysis of PCR products by 2.0% agarose gel electrophoresis
[0036] 5. Interpretation of results
[0037] The result interpretation standard of this kit is as follows Table 2
[0038]
Embodiment 3
[0040] Interpretation of sequencing results and medication guidance
[0041] Using the DNA extracted from the cancer tissue of Example 1 as a template, after amplification with the above-mentioned kit, the sequencing results are obtained, and the corresponding medication instructions are given as follows according to the sequencing results:
[0042]
[0043] The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements and improvements made within the spirit and principles of the present invention should be included in the protection of the present invention. within range.
[0044] sequence listing
[0045] Anhui Anlong Gene Medical Laboratory Co., Ltd.
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