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Primer group and kit

A primer set and kit technology, applied in recombinant DNA technology, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of complicated procedures, long inspection period, time-consuming and laborious amniotic fluid culture method, etc.

Inactive Publication Date: 2018-05-18
HAINAN MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The traditional chromosome inspection method is G-banding karyotype analysis technology, but this technology has great limitations: the amniotic fluid culture method is cumbersome, the inspection cycle is long, time-consuming and laborious; and the amniotic fluid cells are not easy to culture, and the reliability of the test results is even higher. Much depends on the experience of the operator, so it is not conducive to large-scale clinical screening and prenatal diagnosis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1 solution composition, configuration

[0064] (1) According to the primer information published by UCSC (http: / / genome.ucsc.edu / ), primers were synthesized by primer synthesis manufacturers. The forward primer in the primer pair added a fluorescent group at the 5' end, and the reverse primer added a random 7-base primer GTGTCTT at the 5' end. The fluorescent color and primer information are shown in Table 1.

[0065] Table 1: STR primer sequences and fluorescent modifications

[0066]

[0067]

[0068] (2) After obtaining the primer powder synthesized by the primer synthesis manufacturer, centrifuge at 12000rpm for 5min, and use nucleic acid-free ddH 2 O dissolves the primers. According to the primer unit on the primer information sheet, dilute to a primer stock solution with a molar concentration of 100 pmol / μL. Vortex and mix well, then centrifuge at low speed to collect the solution at the bottom of the tube.

[0069] (3) Prepare PCR reaction s...

Embodiment 2

[0075] Embodiment 2PCR operation

[0076] (1) PCR reaction solution A, PCR reaction solution B, enzyme solution, 13-trisomy positive quality control, 18-trisomy positive quality control, 21-trisomy positive quality control, negative quality prepared in Example 1 The control and blank quality control solutions were thawed and vortexed for 5s, then centrifuged at low speed for 10s before use. According to the form of "specimen number + 6" required for the experiment, draw PCR reaction solution A based on 18.8 μL, and put it into 1.5mL EP tube 1; use the same method to draw PCR reaction solution B, and put it into 1.5mL EP tube 2 .

[0077] (2) Pipette 0.2uL of enzyme solution into the above-mentioned EP tube for mixing. Vortex for 5 s, centrifuge at low speed for 10 s and set aside.

[0078] (3) Pipette 19 μL of the mixture containing PCR reaction solution A and enzyme solution into a 0.2mL PCR tube, or 0.2mL 8-strip tube; operate the mixture containing PCR reaction solution ...

Embodiment 3ABI3500

[0082] Example 3 ABI3500 computer operation

[0083] (1) Vortex and mix the PCR product of Example 2 for 5 s, centrifuge at low speed for 10 s, and set aside.

[0084] (2) With the ratio of 9.5 μL HiDi fomamide and 0.5 μL Genescan-500 (LIZ) SIZE STD KIT, mix the solution according to the form of “PCR product number + 1”. Vortex for 5 s and centrifuge at low speed for 10 s.

[0085] (3) Pipette 10 μL of the above solution into a new 0.2mL PCR tube or 0.2mL 8-strip tube, add 1 μL of the product to it at the same time, and cover the tube cap. Vortex for 5 s and centrifuge at low speed for 10 s.

[0086] (4) Put the PCR tube into the PCR instrument, set the program: 94°C for 5 minutes, and immediately transfer to ice for cooling for 5 minutes after the end.

[0087] (5) Using the 96-well plate matched with the ABI Genetic Analyzer, transfer the solution described in (4) into the wells of the 96-well plate one by one.

[0088] (6) Cover with the matching plug of ABI Genetic Ana...

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Abstract

The invention provides a primer group and a kit containing the primer group. According to the embodiment of the invention, the primer group is designed by aiming at 4 STR (short tandem repeat) sites selected by the 13th, the 18th and the 21st chromosomes correspondingly, and can be applied to multiple amplification. The primer group according to the embodiment of the invention can be applied to rapid detection on abnormal number of 21-trisome, 18-trisome and 13-trisome chromosomes.

Description

technical field [0001] The invention relates to the field of biological detection. Specifically, the present invention relates to primer sets and kits. Background technique [0002] Chromosomal disease is a common genetic disease in humans. The incidence rate in surviving newborns is about 1 / 200, and about 80% of them are abnormal chromosome numbers, including trisomy 21, trisomy 18, and trisomy 13. [0003] The traditional chromosome inspection method is G-banding karyotype analysis technology, but this technology has great limitations: the amniotic fluid culture method is cumbersome, the inspection cycle is long, time-consuming and laborious; and the amniotic fluid cells are not easy to culture, and the reliability of the test results is even higher. It largely depends on the experience of the operator, so it is not conducive to large-scale clinical screening and prenatal diagnosis. [0004] Due to the characteristics of prenatal diagnosis, there is an urgent need for a ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/6883
CPCC12Q1/6883C12Q2600/156C12Q2600/166
Inventor 李崎马燕琳陈宏健周繇
Owner HAINAN MEDICAL COLLEGE
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