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Optimizing Method of Extracted Genomic DNA from Fish Blood

Xiaomin Liu

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Abstract

The genomic DNA extracted from frozen whole blood of rainbow trout with methods of chloroform-potassium acetate,rapid genomic DNA extraction(RGDE),improved RGDE,traditional phenol-chloroform and DNA reagent kit respectively,and detected by agarose gel electrophoresis.The results showed that mothode of improved RGDE can obtain better genomic DNA of fish.Then,the effect of extracting with improved RGDE methods were compared by ultraviolet spectrophotometric method with other mothods such as traditional phenol-chloroform from blood or muscle and DNA reagent kit from blood.The results showed that the average concentration of DNA extracted with method improved RGDE was 1 050 ng/μL,and was very significant higher than that by DNA kit or by phenol-chloroform,and the average output of DNA was 1 575 μg/mL,and the rate of A260/A280 was between 1.65~1.97.It only spent 2 hours to extract DNA with the method of improved RGDE,and there were many obvious merits such as quick speed,low cost,no pollution and no using specific installations and so on,and to amplify by random primers,the rusults was stable,and the electrophoresis banding was clear.

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What this paper is about

The genomic DNA extracted from frozen whole blood of rainbow trout with methods of chloroform-potassium acetate,rapid genomic DNA extraction(RGDE),improved RGDE,traditional phenol-chloroform and DNA reagent kit respectively,and detected by agarose gel electrophoresis.The results showed that mothode of improved RGDE can obtain better genomic DNA of fish.Then,the effect of extracting with improved RGDE methods were compared by ultraviolet spectrophotometric method with other mothods such as traditional phenol-chloroform from blood or muscle and DNA reagent kit from blood.The results showed that the average concentration of DNA extracted with method improved RGDE was 1 050 ng/μL,and was very significant higher than that by DNA kit or by phenol-chloroform,and the average output of DNA was 1 575 μg/mL,and the rate of A260/A280 was between 1.65~1.97.It only spent 2 hours to extract DNA with the method of improved RGDE,and there were many obvious merits such as quick speed,low cost,no pollution and no using specific installations and so on,and to amplify by random primers,the rusults was stable,and the electrophoresis banding was clear.

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Available abstract

The genomic DNA extracted from frozen whole blood of rainbow trout with methods of chloroform-potassium acetate,rapid genomic DNA extraction(RGDE),improved RGDE,traditional phenol-chloroform and DNA reagent kit respectively,and detected by agarose gel electrophoresis.The results showed that mothode of improved RGDE can obtain better genomic DNA of fish.Then,the effect of extracting with improved RGDE methods were compared by ultraviolet spectrophotometric method with other mothods such as traditional phenol-chloroform from blood or muscle and DNA reagent kit from blood.The results showed that the average concentration of DNA extracted with method improved RGDE was 1 050 ng/μL,and was very significant higher than that by DNA kit or by phenol-chloroform,and the average output of DNA was 1 575 μg/mL,and the rate of A260/A280 was between 1.65~1.97.It only spent 2 hours to extract DNA with the method of improved RGDE,and there were many obvious merits such as quick speed,low cost,no pollution and no using specific installations and so on,and to amplify by random primers,the rusults was stable,and the electrophoresis banding was clear.

Key concepts: genomic DNA, DNA, Chromatography, Chloroform, Agarose gel electrophoresis, DNA extraction, Chemistry, Agarose

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