A hybridization buffer, a kit for oligonucleotide acgh chips, and a method for detecting variation in chromosome copy number

A hybridization buffer and oligonucleotide technology, applied in the field of molecular biology, can solve problems such as weak hybridization signals, and achieve the effects of improving detection accuracy and improving hybridization leakage.

Active Publication Date: 2021-06-29
CAPITALBIO CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] In view of this, the purpose of the present invention is to address the problems in the prior art, to provide a hybridization buffer, which is suitable for use with aCGH chips modified by high molecular amination three-dimensional polymers, and to improve hybridization leakage and weak hybridization signals. , The problem of uneven hybridization, improve the detection rate of known variant CNV

Method used

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  • A hybridization buffer, a kit for oligonucleotide acgh chips, and a method for detecting variation in chromosome copy number
  • A hybridization buffer, a kit for oligonucleotide acgh chips, and a method for detecting variation in chromosome copy number
  • A hybridization buffer, a kit for oligonucleotide acgh chips, and a method for detecting variation in chromosome copy number

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1, the choice of sample

[0051] In Nigms's human genetic cell library, some cell samples have been purchased and used by chromosomal abnormal detection platforms such as Affymetrix, Illumina, Agilentand Nimblegen. The applicant selects the purchase of the quality control GDNA and control GDNA with representative and known chromosomes abnormalities, and is used in the technical solution of the present invention. Specific sample information is shown in Table 1:

[0052] Table 1 Sample information

[0053]

Embodiment 2

[0054] Example 2, GDNA tags and quality control

[0055] Na04671 and Na12891 were added to 0.2 ml of centrifuge tubes, respectively, followed by Cy3 / Cy5-Random Primer, and completely mixed after fragmentation, 98 ° C and then degenerate it. Ice bath for 2 minutes. Add DNTP, Klenow enzyme mixed 37 ° C for 2 h after mixing, terminate the reaction after the reaction is added, and the labeled product is added to 80% ethanol to dry after drying. Add DDH. 2 O Dissolve. Whether the quality control product and the fluorescent incorption efficiency in accordance with NanoDrop-2000 spectrophotometry are in the labeling efficiency at 13-15 pmol / μg; DNA label synthesis yield is in the range of 25-28 μg, and the results are within the moisturization range.

Embodiment 3

[0056] Example 3, chip hybridization and scanning

[0057] After the labeling material is passed, the Na04671 and Na12891 samples were taken at each 12 μg of labeling product, and 3 × buffer Mix 15.0 μl, COT-1DNA 5.0 μL, 1 μl of formamide, 2.25 μl of dodecyl sulfate, 98 EtOAc EtOAc. The hybrid mixed liquid is added to the surface of the chip, and the surface of the chip is laminated under the action of the hybrid cover, and the solution cavity is formed with the hybrid cover, and placed in Biomixer TM II hybrids overnight overnight for 14 hours.

[0058] 3 × buffer Mix:

[0059]

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Abstract

The invention belongs to the field of molecular biology and provides a hybridization buffer solution, a reagent kit of an oligonucleotide comparison genome hybridization chip and a method for detecting variation in chromosome copy number. The hybridization buffer of the present invention is composed of buffer Mix, formamide, lithium dodecyl sulfate and Cot-1DNA, which can significantly improve the problems of hybridization leakage, weak hybridization signal, uneven hybridization, and poor specificity, and improve the known The detection accuracy of variant CNV.

Description

Technical field [0001] The present invention belongs to the field of molecular biology, and more particularly to a hybridization buffer, an oligonucleotide comparative genomic hybridized chip (oligonucleotide ACGH chip) kit, and a method of detecting the number of chromosome copies. Background technique [0002] The abnormal chromosomal abnormalities accounted for more than 50% of spontaneous abortion, dead, early death, and the incidence of newborn is about 1%, which is an important reason for sexual development and male and female, and infertility. It is also a congenital heart One of the important causes of illness, intelligent development. With the development of chromosomal straps, PCR technology, DNA testing technology, etc., the understanding of chromosome distortion and disease relationship is increasing, and the dye body disease is increasing. Comprehensive information on many countries, about 15% of pregnancy abortion, which is caused by abnormal chromosomes, namely 5% ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6837C12Q1/6883
CPCC12Q1/6837C12Q1/6883
Inventor 王亚辉郭弘妍王辉邢婉丽程京单万水刘厚明
Owner CAPITALBIO CORP
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