Primer for inhibiting sequence-specific amplification of 18S rDNA of shrimps and oysters and application thereof

A specific, oyster technology, applied in the field of PCR primers, to achieve the effect of convenient research

Active Publication Date: 2018-06-01
广州鸿真生物科技有限公司
View PDF8 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no research report on the primers used to inhibit the

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
  • Primer for inhibiting sequence-specific amplification of 18S rDNA of shrimps and oysters and application thereof
  • Primer for inhibiting sequence-specific amplification of 18S rDNA of shrimps and oysters and application thereof
  • Primer for inhibiting sequence-specific amplification of 18S rDNA of shrimps and oysters and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Design of inhibitory primers

[0024] Suppressing primer design scheme:

[0025] 1. Design of C3 spacer

[0026] There is a certain overlap between the 5'end (18-25nt) and the F(R) 3'end of the universal primer. The subsequent sequence should be as consistent as possible with the 18S gene sequence to be suppressed, and different from the gene sequence that is not desired to be suppressed, and finally at the end Add C3spacer.

[0027] 2. Design of dual-priming oligonucleotides

[0028] The design of the 5'end is similar to the 5'end of C3, except that 44444 (4=dlnosine) is added before the 3'end (6-12nt), and finally C3spacer is added to the end.

[0029] The design process of suppression primers:

[0030] Such as figure 1 As shown, according to Penaeus vannamei (GenBank accession no.EU920969.1), Penaeus vannamei (GenBank accession no.EU118282.1), Penaeus vannamei (GenBank accession no.AF186250.1), Penaeus vannamei (GenBank accession no. AF124597.1), Penaeus vannamei (Ge...

Embodiment 2

[0035] Example 2 Verification of suppression primers

[0036] The DNA of Penaeus vannamei and Hong Kong oyster were extracted using the Mollusk DNA Extraction Kit (Magen, Suzhou, China), and the DNA of plankton microorganisms and zooplankton in the water filtered onto the membrane were extracted using the Water DNA Extraction Kit (Magen, Suzhou, China) ) The specific operation is carried out in accordance with the instructions on the manual.

[0037] For the inhibitory primer nmb-BP2-DPO, configure the inhibition reaction system with different concentrations of 0.01μM, 0.02μM, 0.04μM, and use ddH 2 O was used as a negative control, and the reaction without inhibitory primers was used as a positive control. The templates used the digestive tract tissue DNA of Penaeus vannamei, the digestive tract tissue DNA of Hong Kong oyster, the planktonic microorganism DNA filtered to the membrane, and the zooplankton DNA. Set up 3 test replicates for different template DNAs, and amplify them by...

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 discloses a primer for inhibiting sequence-specific amplification of 18S rDNA of shrimps and oysters. The primer mainly comprises base sequences as shown in SEQ ID NO.1, five deoxyhypoxanthines inserted among the base sequences AG, and C3 spacers at the tail ends of the base sequences. The primer can effectively inhibit the amplification of host genes and enhance the amplification effect of 18S rDNA of other eukaryotes (such as protozoa, algae and fungi) in host bodies. The invention further discloses the application of the primer in preparing a reagent with the effect of inhibiting the sequence-specific amplification of 18S rDNA of the shrimps and oysters.

Description

Technical field [0001] The invention belongs to the technical field of PCR primers, and specifically relates to a primer for inhibiting the specific amplification of 18S rDNA sequences of shrimp and oysters and its application. Background technique [0002] The genome of eukaryotic animal hosts (such as oysters and shrimps) contains 18S rDNA gene sequences. At the same time, its body (such as the intestine) also carries a variety of eukaryotic microorganisms (such as undigested algae food, endosymbiotic fungi, parasites), and their genomes also contain 18S gene sequences. The 18S sequences between different species are Certain similarity. [0003] In order to study the eukaryotic microorganisms present in the host, people usually use the PCR method to specifically amplify the 18S gene with universal primers. However, due to the inevitable interference of the host's own 18S gene sequence, the host's 18S sequence in the amplified product occupies most of the proportion, which affec...

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): C12N15/11C12N15/10
CPCC12N15/10C12N15/113
Inventor 姜敬哲戚认杰刘聪翟立广
Owner 广州鸿真生物科技有限公司
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

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap