Improved reverse dot hybridization method and application thereof

A technology of reverse dot hybridization and hybridization solution, which is applied in the application field of reverse dot hybridization method in products, can solve the problems of long operation time, many operation steps, and high price of automatic hybridization instrument, and achieve the goal of reducing steps and time Effect

Active Publication Date: 2011-06-22
DAAN GENE CO LTD
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, the entire hybridization process of RDB technology requires many steps, and the operation time is relatively long, which has become a resistance to large-scale promotion and application.
Based on this demand, although corresponding automated hybridization instruments have appeared, the price of automated hybridization instruments is expensive, which is

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Improved reverse dot hybridization method and application thereof
  • Improved reverse dot hybridization method and application thereof
  • Improved reverse dot hybridization method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0033] Example 1 Comparison between the traditional method and the method of the present invention in the detection of thalassemia

[0034] The beta thalassemia detection kit produced by Daan Gene Co., Ltd. of Sun Yat-sen University is used to compare the detection results of the method of the present invention and the kit method (ie, the traditional method).

[0035]

[0036]

[0037] The enzyme used in method A is horseradish peroxidase, and the enzyme used in method B is alkaline phosphatase. Three clinical samples were selected, namely CD41 / 42 heterozygotes, IVS2-654 heterozygotes, and β globin were normal. From the experimental results, method A, method B and kit method can all correctly detect the corresponding results, and the three are implemented There is no difference in effect (see attached figure 1 Numbers 1 to 3 are traditional methods, 4 to 6 are methods A of the present invention, and 7-9 are methods B) of the present invention. The method of the present invention ha...

Example Embodiment

[0038] Example 2 Application of the method of the present invention in HCV genotyping detection

[0039] 1. The primer and probe sequences used in the kit are as follows:

[0040] Hepatitis C virus (HCV) nucleic acid (RNA) reverse transcription primer sequence is:

[0041] Reverse transcription (RT) primer: 5'-GCTCATGGTGCACGGTCTACGAGACCT-3' (SEQ ID NO: 1)

[0042] The upstream and downstream primer sequences used in the polymerase chain reaction amplification system are:

[0043] PCR upstream primer: 5'-TCTAGCCATGGCGTTAGTATGAGTGT-3' (SEQ ID NO: 2)

[0044] PCR downstream primer: 5'-CACTCGCAAGCACCCTATCAGGCAGT-3' (SEQ ID NO: 3)

[0045] The oligonucleotide probes used in the hybridization reaction are all labeled with an amino group at the 5'end, and the probe IC is labeled with an amino group at the 5'end and biotin at the 3'end. The sequences are:

[0046] Probe 1: 5'-CAATGCCTGGAGATTTGGGCG-3' (SEQ ID NO: 4)

[0047] Probe 1b-1: 5'-CCGCGAGACTGCTAGCCG-3' (SEQ ID NO: 5)

[0048] Probe 1b-2: 5'-...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to an improved reverse dot hybridization method. By adopting the method, the steps in the conventional reverse dot hybridization method are optimized, so that the steps and time of reverse dot hybridization experiment operation are reduced without causing any difference on a detection result. The application of the method to a reverse dot hybridization product contributes to popularization of the method in the field of clinical detection.

Description

technical field [0001] The invention relates to an improved reverse dot hybridization method and the application of the improved reverse dot hybridization method to products. Background technique [0002] In 1989, reverse dot blot (RDB) technology was proposed by Saiki et al. (Saiki RK, et al.Proc Natl Acad Sci USA.1989, 86 (16): 6230-4), which was based on PCR- A practical improvement is made on the basis of ASO (Polymerase chain reaction&allelespecific oligonucleotide) technology. Henry Erlich, Randall Saiki and their colleagues proposed an effective improvement in the detection of HLA. This method is to label the target gene with radioactive elements during the PCR process, and then hybridize with the nylon membrane to detect the mutation of the target gene. This method can detect multiple target genes under the same conditions, thus greatly improving the detection efficiency. Subsequently, other scientists confirmed that the method can distinguish mutations of single b...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68G01N21/75
Inventor 程钢林炳生张帆陈华云
Owner DAAN GENE CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products