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Simple repetition sequence primer for moerella iridescens

A simple repeating sequence, rainbow technology, applied in recombinant DNA technology, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of complex operating conditions, low annealing temperature, high cost, and reduce enzyme digestion steps, The effect of high annealing temperature and low cost

Inactive Publication Date: 2012-08-08
HUAIHAI INST OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a simple repeat sequence primer for the rainbow clam, which can overcome the short primers in the prior art, low annealing temperature, complicated operating conditions, and low cost. high defect

Method used

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  • Simple repetition sequence primer for moerella iridescens
  • Simple repetition sequence primer for moerella iridescens
  • Simple repetition sequence primer for moerella iridescens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. The simple repeat sequence primer of the rainbow clam, which is a primer set composed of the following 10 primer sequences:

[0038]

Embodiment 2

[0039] Example 2. The application of the simple repeat sequence primer of rainbow clam, its steps are as follows,

[0040] 1.10 Synthesis of standard primers

[0041] 10 primers in the table of Example 1 were synthesized by a bioengineering company, each 2 OD.

[0042] 2. Extraction of the genome of the rainbow clam

[0043] Collect 30 samples of wild populations of Rainbow clams (the number of populations is not limited, but at least two), and bring fresh samples back to the laboratory, and use the standard phenol-chloroform method to extract genomic DNA. Specific process and reagent formula, the genomic DNA of rainbow clam adopts SDS lysate formula: 10mmol / LTris.Cl (pH=8.0), 0.1mmol / L EDTA (pH=8.0), 0.5%SDS, the step of extraction is to take Put 50mg of muscle tissue of Rainbow clam, grind it into fine powder in liquid nitrogen, put it into a 2.0ml centrifuge tube, add 600μl of SDS tissue lysate, put it in a water bath shaker at 55°C, and add 10μl of proteinase K after ly...

Embodiment 3

[0053] Example 3. The application of the simple repeat sequence primer of rainbow clam, its steps are as follows,

[0054] 1. Extract the genomic DNA of the breeding line of P.

[0055] (1) SDS lysate formulation: 10 mmol / L Tris.Cl (pH=8.0), 0.1 mmol / L LEDTA (pH=8.0), 0.5% SDS.

[0056] (2) Step: Take 50 mg of muscle tissue from the legs of the rainbow clam, grind it into a fine powder in liquid nitrogen, put it into a 2.0ml centrifuge tube, add 600 μl of SDS tissue lysate, and put it in 55°C Water bath shaker, after 1 hour of lysis, add 10 μl of proteinase K (20 mg / ml), continue lysis for 3 to 4 hours, then take out the centrifuge tube and flick the bottom of the centrifuge tube with your fingers, let the unlysed tissue fully contact with the lysate, and make the tissue Cells can be fully lysed;

[0057] After lysis, add 600 μl Tris saturated phenol to each centrifuge tube, shake gently, invert 45-55 times, and let stand at room temperature for 10-15 minutes;

[0058] 120...

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Abstract

The invention relates to a simple repetitive sequence primer for Rainbow Moerella, which is characterized in that the simple repetitive sequence primer for the Rainbow Moerella is a primer group which is formed by 10 Rainbow Moerella primers. The screened amplified products of the primer are clear and stable, have good polymorphism, can be applied in idioplasm evaluation and idioplasm identification of the Rainbow Moerella, and diversity evaluation of colonies and remains of the Rainbow Moerella, and have the characteristics of quickness, economy, accuracy, simple and convenient operation andso on.

Description

technical field [0001] The invention relates to a set of primers for germplasm identification and evaluation and genetic structure analysis, in particular to a set of simple repetitive sequence primers for germplasm identification and evaluation and genetic structure analysis of clams. Background technique [0002] At present, the molecular markers commonly used to evaluate biological genetic diversity and germplasm identification have a certain degree of defects, such as RAPD technology (Random amplified polymorphic DNA, Random amplified polymorphic DNA), often because RAPD primers are too short by 10 bases The base and annealing temperature are too low, generally 36-37°C, leading to some non-specific results of the amplified product, and its reliability and authenticity are the main defects; and AFLP technology (amplified fragment length polymorphism, Amplified fragment length polymorphism), However, its wide application is limited by complex operating conditions such as e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 董志国王美珍李晓英陈汉春孟学平程汉良阎斌伦
Owner HUAIHAI INST OF TECH
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