Kit for detecting human BDNFmet gene variation

A gene mutation and kit technology, applied in the biological field, can solve problems such as unsuitable for large sample size screening and expensive sequencing technology

Inactive Publication Date: 2009-12-09
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Restriction enzyme digestion methods often have incomplete digestion, which affects the accuracy

Method used

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  • Kit for detecting human BDNFmet gene variation
  • Kit for detecting human BDNFmet gene variation
  • Kit for detecting human BDNFmet gene variation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0037] 1. Take 3ml of peripheral anticoagulant blood, centrifuge horizontally at 3000rpm for 10min, absorb 300ul buffy coat into a 1.5ml EP tube, and store it at -80°C for later use.

[0038] 2. Add 700ul sterile distilled water to the leukocytes, invert and mix well, centrifuge at 10000rpm for 3min, and discard the supernatant.

[0039] 3. Add 80ul Kl (5mol / L) to the precipitate, shake for 30s, add 300ul normal saline and 450ul chloroform-isoamyl alcohol (24:1), shake vigorously for 20s, and centrifuge at 12000rpm for 5min.

[0040] 4. Take about 400ul of the upper aqueous phase into a new EP tube, add 180ul of isopropanol, invert and mix until flocculent precipitation appears.

[0041] 5. Centrifuge at 12000rpm for 5min, discard the supernatant, add 900ul of 70% ethanol, shake to float the precipitate.

[0042] 6. Centrifuge at 12000rpm for 5min, discard the supernatant, and air dry for about 5min. Add 100ul TE to dissolve the DNA to obtain the sample DNA to be tested, and...

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Abstract

The invention relates to a kit used for accurately detecting whether valine at the 66th position of BDNF in a specimen is variated into methionine by a fluorescence PCR detection technique. The invention provides effective guide for clinically and individually treating mental sickness, such as anxiety neurosis, melancholia, post-traumatic stress syndrome, and the like correlative to BDNF gene variation.

Description

technical field [0001] The invention belongs to the field of biotechnology, and is a kit for accurately detecting whether valine at position 66 of brain-derived neurotrophic factor (BDNF) in a sample is mutated into methionine through fluorescent PCR detection technology. Background technique [0002] Traditional detection techniques for detecting whether the valine at position 66 of brain-derived neurotrophic factor (BDNF) in a specimen is mutated into methionine include: PCR-restriction enzyme digestion technology and PCR-sequencing technology. Both of these methods require subsequent testing after PCR testing. Restriction enzyme digestion methods often have incomplete digestion, which affects the accuracy of detection; and sequencing technology is expensive and not suitable for large-scale screening. The present invention does not need follow-up detection after PCR, is quick and easy; detects with a probe, and the result is accurate; the cost is low, and it is especially...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64
Inventor 陈哲宇姜虹王越于卉
Owner SHANDONG UNIV
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