Probe and method for carrying out hybrid capturing on gene duplication area

A technology of gene duplication and hybridization capture, applied in biochemical equipment and methods, recombinant DNA technology, microbial assay/inspection, etc. Effect

Inactive Publication Date: 2019-05-21
天津诺禾医学检验所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a probe and method for hybridization and capture of gene repeating regions, so as to solve the technical problem that the probe design and effective capture of high-repetition regions cannot be carried out in the existing hybridization and capture methods

Method used

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  • Probe and method for carrying out hybrid capturing on gene duplication area
  • Probe and method for carrying out hybrid capturing on gene duplication area
  • Probe and method for carrying out hybrid capturing on gene duplication area

Examples

Experimental program
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Effect test

Embodiment 1

[0055] Sample information: cell line H1299DNA

[0056] Reagents and instruments are shown in Table 3 below:

[0057] table 3

[0058] equipment name

manufacturers

model

Savant TM DNA SpeedVac TM concentrator

Thermo

DNA120-230

thermal cycler

BIO-RAD

T100 TM Thermal Cycler

96-well magnetic stand

Invitrogen

12331D

Vortex shaker

Dinghaoyuan Technology

VS-1

mini smart hand centrifuge 6×1.5mL

Dinghaoyuan Technology

0401061

mini smart hand centrifuge 2×8×0.2mL

Dinghaoyuan Technology

0401062

Magnetic stand 1.5-2mL

Dry Thermostat

Hangzhou Aosheng Instrument Co., Ltd.

K30

Reagent

manufacturers

Item No.

10× Wash Buffer Ⅰ

IDT

1072273

10× Wash Buffer II

IDT

1072274

10× Wash Buffer III

IDT

1072275

10× Enhanced Cleaning Fluid

IDT

1072276

2× hybridization buffer

IDT

107227...

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Abstract

The invention discloses a probe and method for carrying out hybrid capturing on a gene duplication area. The probe is a primer probe with a conserved area beside a target gene duplication area or a specific position in the duplication area as a 3'terminal and with a detecting mark. According to the technical scheme, based on the primer amplification and extension theory, the specificity requirement of primer extension on a sequence, especially a 3' terminal sequence is larger than the specificity requirement of base complementary pairing in hybrid capturing for the sequence, the conserved areabeside the gene duplication area or a relatively specific position in the duplication area is adopted as the 3'terminal with the highest requirement for the specificity of the primer, the primer probe with the detecting mark is designed as the capturing probe, and in combination with a traditional liquid phase hybrid capturing system, the capturing efficiency of the gene duplication area is specifically improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a probe and method for hybridizing and capturing gene repeating regions. Background technique [0002] The liquid-phase hybridization capture technology is developed according to the principle of base complementarity, through the direct hybridization of the target DNA fragment and the biotin-labeled probe in the solution, and then the target DNA fragment is anchored by the reaction of biotin avidin fixed on magnetic beads with avidin. Magnetic beads are covalently combined with high-purity streptavidin and magnetic particles, which can be used to capture biotin-labeled substrates. The biotin-streptavidin interaction is strong and the non-specific binding rate is very low. The captured substrate can meet the requirements of subsequent experiments. The non-target DNA is washed away by washing the magnetic beads, and then the DNA of the target region is obtained by eluting the magneti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6813C12N15/11
Inventor 梁永王棪李雷成岗李瑞强
Owner 天津诺禾医学检验所有限公司
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