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RT-PCR primer and method for amplifying estrogen receptor alpha genes in female chinemys reevesii

An estrogen receptor, RT-PCR technology, applied in the field of molecular biology research, to achieve the effect of research and cost saving

Inactive Publication Date: 2018-02-02
ANHUI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there has been no related research on the estrogen receptor α gene in female tortoises in previous reports.

Method used

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  • RT-PCR primer and method for amplifying estrogen receptor alpha genes in female chinemys reevesii
  • RT-PCR primer and method for amplifying estrogen receptor alpha genes in female chinemys reevesii
  • RT-PCR primer and method for amplifying estrogen receptor alpha genes in female chinemys reevesii

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1) Extraction of total RNA: Using rapid hot phenol extraction method, take 100mg of tubal vaginal tissue samples, put them into a grinder and grind them thoroughly, then place them in a sterilized 2.0mL EP tube; in the EP tube Add 1 mL of Trizol solution, incubate at room temperature for 5 min to fully lyse; centrifuge at 12,000 g for 10 min at 4°C, take out the upper aqueous phase and transfer it to another 2.0 mL EP tube; add 0.2 mL of chloroform (dichloromethane) to the EP tube ) and 1mL Trizol solution, shake gently for 15s, and place at room temperature for 2min; then centrifuge at 12000g and 4°C for 15min, take out the upper aqueous phase and transfer it to another 2.0mL EP tube; add 0.5ml isotropic acid to the EP tube Propanol and 1mL Trizol, mix well, incubate at room temperature for 10min; centrifuge at 12000g, centrifuge at 4°C for 10min, discard the supernatant; add 1mL of 75% ethanol to wash (just invert the EP tube up and down gently); centrifuge at 7500g, r...

Embodiment 2

[0039] Prepare according to the preparation method of Example 1, the difference is that the tubal vaginal part in Example 1 is replaced by the tubal uterine part, and the electrophoresis results are as follows: figure 1 shown. (lane number is 2)

Embodiment 3

[0041] Prepare according to the preparation method of Example 1, the difference is that the oviduct vaginal part in Example 1 is replaced by the isthmus of the oviduct, and the electrophoresis results are as follows figure 1 shown. (lane number is 3)

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Abstract

The invention discloses a RT-PCR primer and method for amplifying estrogen receptor alpha genes in female chinemys reevesii. The RT-PCR primer comprises a primer pairs as shown in SEQ ID No:1 and SEQID No:2. The method comprises the following steps: 1) extracting total RNA of a to-be-amplified sample; 2) reversely transcribing the extracted total RNA into cDNA; 3) amplifying the cDNA by using theRT-PCR primer; PT-PCR primer is the PT-PCR primer. According to the invention, a set of RT-PCR primers are designed, the total RNA of the to-be-amplified sample is extracted, and then reverse transcription is conducted to obtain the cDNA. The cDNA is amplified by using the RT-PCR primer. Under the premise of cost reducing, a great number of estrogen receptor alpha genes in female chinemys reevesii fallopian tubes are amplified rapidly and accurately, and large-scale and more intuitive studies of the estrogen receptor alpha genes in the female chinemys reevesii are effectively achieved.

Description

technical field [0001] The invention relates to the field of molecular biology research, in particular to RT-PCR primers and methods for amplifying the estrogen receptor alpha gene of female tortoises. Background technique [0002] Turtles (Chinemys reevesii) belong to the class Reptiles, Turtles, Freshwater Turtles, and Turtles. Turtles have high medicinal and nutritional value and are deeply loved by people. In recent decades, human economic activities have intensified, and the ecological environment on which turtles depend has been severely damaged. In addition, overfishing of wild turtles, habitat destruction, chemical pollution, and river dam building have led to fragmentation and islanding of their living waters. , has caused a sharp decline in the number of wild tortoises, and wild species are in danger of extinction. It can be seen that the protection of wild populations of tortoises is imminent. With the increasing scarcity of wild populations, the scale of artif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/124C12Q2600/158C12Q2521/107
Inventor 卜兴江陆文康何可欣王湘林
Owner ANHUI NORMAL UNIV
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