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New short nucleotide tandem repeat site and application thereof

A short tandem repeat and nucleotide sequence technology, applied in the field of short nucleotide tandem repeat sequence sites, can solve the problems of poor compatibility and achieve high compatibility, high heterozygosity, and high resolution

Active Publication Date: 2014-10-15
GENESKY DIAGNOSTICS SUZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Therefore, in order to overcome defects such as poor compatibility, presence of CNV or location in highly repetitive sequence regions, a new type of short tandem repeat sequence with high discrimination is still needed in this field

Method used

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  • New short nucleotide tandem repeat site and application thereof
  • New short nucleotide tandem repeat site and application thereof
  • New short nucleotide tandem repeat site and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0154] Combining the new STR locus G5S0001 of the present invention with 10 commonly used loci (VWA, D16S539, D5S818, D7S820, D13S317, D8S1179, D18S51, TH01, D2S1338, AMELO), multiplex fluorescent PCR technology was used to check the samples, and the four used The samples were three samples from a related family and one unrelated control sample. The primer sequences of G5S0001 and 10 commonly used loci are shown in Table 2.

[0155] Table 2 Primer sequences

[0156]

[0157] Note: PET, VIC, NED, FAM four fluorescent dyes ((Applied Biosystems, USA company).

[0158]

[0159] The specific experimental steps are as follows:

[0160] 1) DNA sample preparation

[0161] Collect blood from 4 selected sample individuals; extract DNA samples through DNA extraction kit and take 1 μl each, perform quality inspection and concentration estimation on the samples by 1% agarose electrophoresis, and then dilute the samples to the working concentration according to the estimated concentr...

Embodiment 2

[0175] Randomly select 20 groups of related and unrelated samples, and use the method of Example 1 for detection, the difference is that only the new STR locus G5S0001 of the present invention is used.

[0176]After testing, after determining the paternal sample, determine the offspring samples that are related to the male parent in the remaining 19 groups of samples. The new STR locus G5S0001 of the present invention can accurately exclude 9 groups of unrelated samples. A higher degree of discrimination was obtained.

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Abstract

The invention discloses a new short nucleotide tandem repeat site and application thereof. A short tandem repeat locus G5S0001 has a sequence shown as SEQ ID NO.1, and can be used for preparation of (a) reagents or kits for genetic relationship analysis; (b) reagents or kits for individual identification; (c) reagents or kits for paternity testing or blood relationship analysis; (d) reagents or kits for detecting whether maternal blood contamination exists in extracted amniotic fluid; and / or (e) kits for detecting whether leucocytes in a bone marrow transplantation receptor are substituted by donor cells. The short tandem repeat locus G5S0001 has high discrimination, and can effectively analyze the genetic relationship.

Description

[0001] technical field [0002] The invention relates to a short nucleotide tandem repeat sequence site and its application. [0003] Background technique [0004] Short tandem repeat (Short Tandem Repeat. STR), also known as microsatellite DNA (microsatellite DNA), is a kind of DNA tandem repeat sequence with a repeat unit of 2-6bp, a repeat number of 10-60, and a fragment length of less than 400bp; The reason is slippage during DNA replication, or base mismatch between the slipping strand and the complementary strand during replication and repair, resulting in the deletion or insertion of one or several repeat units. STR has a wide distribution in the genome, multiple alleles, high heterozygosity, and stable typing results. STR inheritance conforms to Mendelian inheritance laws (Koreth, J., et al. 1996. Microsatellites and PCR ge-nomic analysis.J Pathol.178: 239-248.) and other characteristics, and when multiple STR loci are jointly detected, the individual recognition a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 姜正文马瑞晓张希
Owner GENESKY DIAGNOSTICS SUZHOU
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