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Kit and application method for combined detection of duck igf-I and igf-IR gene mrna expression

A technology of IGF-IR and IGF-I, which is applied in the fields of biochemical equipment and methods, microbial determination/inspection, bioreactor/fermenter combination, etc. The problem of low level and high level, to achieve the effect of low price

Inactive Publication Date: 2015-08-19
朱文奇
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] my country is the world's largest producer and consumer of waterfowl, but the overall production level of my country's waterfowl industry is not high, the competitiveness is not strong, the technology in the field of waterfowl is very weak, and there are few scientific research results available, especially related to duck muscle. There are very few research results on growth and development molecular technology. Therefore, in order to quickly and effectively improve the production level of my country's duck industry in a short period of time, it is necessary to use advanced molecular biology techniques and achievements to accelerate the development of my country's duck industry.

Method used

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  • Kit and application method for combined detection of duck igf-I and igf-IR gene mrna expression
  • Kit and application method for combined detection of duck igf-I and igf-IR gene mrna expression
  • Kit and application method for combined detection of duck igf-I and igf-IR gene mrna expression

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Embodiment 1

[0070] Example 1, using the kit and method provided by the present invention to conduct relative quantitative analysis on the liver tissue of 4-week-old Gaoyou duck.

[0071] 1. Sample and RNA extraction

[0072] Five liver tissues of 4-week-old Gaoyou ducks were collected. Liver total RNA extraction by TRNzol-A + Total RNA extraction kit (DP421) instructions. RNA samples were detected by 1.4% agarose-formaldehyde denaturing gel electrophoresis and UV spectrophotometer, OD 260／280 1.8 to 2.0, to ensure reliable quality of RNA samples, and normalize the RNA concentration to 1 μg / μl.

[0073] 2. Primer design

[0074] Primer design was performed according to step 1 of the method. IGF-I gene chimeric specific upstream and downstream primers (P1 and P2), IGF-I R gene chimeric specific upstream and downstream primers (P3 and P4), reference gene ACTB gene chimeric specific upstream and downstream primers (P5 and P6).

[0075] The information expressed by the IGF-I gene chimeri...

Embodiment 2

[0092] Example 2, using the kit and method provided by the present invention to carry out relative quantitative analysis on the liver tissue of 2-week-old Jinding duck.

[0093] 1. Sample and RNA extraction

[0094] Three liver tissues of 2-week-old Jinding ducks were collected. Liver total RNA extraction by TRNzol-A + Total RNA extraction kit (DP421) instructions. RNA samples were detected by 1.4% agarose-formaldehyde denaturing gel electrophoresis and UV spectrophotometer, OD 260／280 1.8 to 2.0, to ensure reliable quality of RNA samples, and normalize the RNA concentration to 1 μg / μl.

[0095] 2. Primer design

[0096] Primer design was performed according to step 1 of the method. IGF-I gene chimeric specific upstream and downstream primers (P1 and P2), IGF-I R gene chimeric specific upstream and downstream primers (P3 and P4), reference gene ACTB gene chimeric specific upstream and downstream primers (P5 and P6).

[0097] The information expressed by the IGF-I gene ch...

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Abstract

The invention discloses a kit for joint detection on mRNA (messenger Ribose Nucleic Acid) expression quantity of IGF-I (Insulin-like Growth Factor) and IGF-IR (Insulin-like Growth Factor I Receptor) genes of ducks and a using method of the kit. The kit comprises a kit body (1), an isolating pad (2), an IGF-I gene mosaic specific upstream and downstream primer (3), an IGF-IR gene mosaic specific upstream and downstream primer (4), a reference gene ACTB (actin beta) gene mosaic specific upstream and downstream primer (5), an IGF-I gene internal control DNA (6), an IGF-IR gene internal control DNA (7) and a Tag enzyme DNA ligase (8). The kit, which is special in design and is stable and efficient, is combined with an MCF-PCR (Malignant Catarrhal Fever-Polymerase Chain Reaction) technology, and thus a detection method is reliable and convenient and is low in price.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a kit for joint detection of duck IGF-I and IGF-I R gene mRNA expression levels and a use method. Background technique [0002] my country is the world's largest producer and consumer of waterfowl, but the overall production level of my country's waterfowl industry is not high, the competitiveness is not strong, the technology in the field of waterfowl is very weak, and there are few available scientific research results, especially related to duck muscle. There are very few research results on growth and development molecular technology. Therefore, in order to quickly and effectively improve the production level of my country's duck industry in a short period of time, it is necessary to accelerate the development of my country's duck industry with the help of advanced molecular biology technology and achievements. [0003] MCF-PCR technology is an economical, high-through...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/00C12Q1/68
CPCC12Q1/686C12Q2537/161C12Q2531/113
Inventor 李慧芳朱文奇张静宋卫涛胡艳宋迟束婧婷朱春红陶志云徐文娟善艳菊
Owner 朱文奇
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