[Objective] This study aimed to investigate a reliable method for DNA ex- traction from Wusuli raccoon dog＇s hair. [Method] Several DNA extraction methods were used to extract DNA from Wusuli raccoon dog hair, includ...[Objective] This study aimed to investigate a reliable method for DNA ex- traction from Wusuli raccoon dog＇s hair. [Method] Several DNA extraction methods were used to extract DNA from Wusuli raccoon dog hair, including Chelex-100 method, PCR buffer method, organic phenol-chloroform method and centrifugal col- umn type kit method. The extracted DNA was analyzed by using PCR amplification and electrophoresis to compare these four DNA extraction methods. [Result] Accord- ing to the results of spectrophotometer detection and gel electrophoresis, nucleic acid extracted by Chetex-100 method had proteins and other impurities; nucleic acid ex- tracted by PCR buffer method was low in concentration; however, DNA extracted by organic phenol-chloroform method and centrifugal column type kit was high in con- centration with no impurity band. [Conclusion] This study had laid the strong founda- tion of scientific theory to further explore the efficient and simple method for extracting DNA from Wusuli raccoon dog hair follicle.展开更多
This study aimed to purify and determine antioxidant activities of different fractions obtained during the purification process of phycocyanin from Spirulina platensis. The dried powder of Spirulina platensis, after g...This study aimed to purify and determine antioxidant activities of different fractions obtained during the purification process of phycocyanin from Spirulina platensis. The dried powder of Spirulina platensis, after ground with sands, was extracted with 50 mM sodium phosphate buffer at pH 6.8 before centrifuged to precipitate unwanted proteins. Then the supernatant was separated by celit column to obtain semi-pure phycocyanin and further purified by treated with ammonium sulfate. The purity of phycocyanin was monitored by measuring the absorbance spectrum from 200 to 700 nm. Its purity ratio A620A280 was determined. The antioxidant activities of the obtained phycocyanin were determined by 2,2＇-azino-bis（3-ethylbenzthiazoline-6-sulphonic acid） （ABTS） assay and lipid peroxidation （linoleie acid） assay. The purity ratio of phycocyanin in the Spirulina crude extract was 0.36 and increased to 2.68 after purification. The fraction with the highest purity ratio of phycocyanin demonstrated the hightest antioxidant activities. For ABTS assay, it presented the Vitamin C Equivalent Antioxidant Capacity （VCEAC） value of 0.0405 ±0.0002 mg of ascorbic acid/mg of sample and the Trolox Equivalent Antioxidant Capacity （TEAC） value of 0.0485 ±0.0002 mg oftrolox/mg of sample respectively. The result from lipid peroxidation assay exhibited IC50 value of 5.9336 ±0.2565 mg/mL. The purification of Spirulinaplatensis crude extract obtained from this study increased the purity ratio of phycocyanin and its antioxidant activities. This will be further investigated for the development into anti-aging cosmetic products.展开更多
基金Supported by National Natural Science Foundation of China (31072018)~~
文摘[Objective] This study aimed to investigate a reliable method for DNA ex- traction from Wusuli raccoon dog＇s hair. [Method] Several DNA extraction methods were used to extract DNA from Wusuli raccoon dog hair, including Chelex-100 method, PCR buffer method, organic phenol-chloroform method and centrifugal col- umn type kit method. The extracted DNA was analyzed by using PCR amplification and electrophoresis to compare these four DNA extraction methods. [Result] Accord- ing to the results of spectrophotometer detection and gel electrophoresis, nucleic acid extracted by Chetex-100 method had proteins and other impurities; nucleic acid ex- tracted by PCR buffer method was low in concentration; however, DNA extracted by organic phenol-chloroform method and centrifugal column type kit was high in con- centration with no impurity band. [Conclusion] This study had laid the strong founda- tion of scientific theory to further explore the efficient and simple method for extracting DNA from Wusuli raccoon dog hair follicle.
文摘This study aimed to purify and determine antioxidant activities of different fractions obtained during the purification process of phycocyanin from Spirulina platensis. The dried powder of Spirulina platensis, after ground with sands, was extracted with 50 mM sodium phosphate buffer at pH 6.8 before centrifuged to precipitate unwanted proteins. Then the supernatant was separated by celit column to obtain semi-pure phycocyanin and further purified by treated with ammonium sulfate. The purity of phycocyanin was monitored by measuring the absorbance spectrum from 200 to 700 nm. Its purity ratio A620A280 was determined. The antioxidant activities of the obtained phycocyanin were determined by 2,2＇-azino-bis（3-ethylbenzthiazoline-6-sulphonic acid） （ABTS） assay and lipid peroxidation （linoleie acid） assay. The purity ratio of phycocyanin in the Spirulina crude extract was 0.36 and increased to 2.68 after purification. The fraction with the highest purity ratio of phycocyanin demonstrated the hightest antioxidant activities. For ABTS assay, it presented the Vitamin C Equivalent Antioxidant Capacity （VCEAC） value of 0.0405 ±0.0002 mg of ascorbic acid/mg of sample and the Trolox Equivalent Antioxidant Capacity （TEAC） value of 0.0485 ±0.0002 mg oftrolox/mg of sample respectively. The result from lipid peroxidation assay exhibited IC50 value of 5.9336 ±0.2565 mg/mL. The purification of Spirulinaplatensis crude extract obtained from this study increased the purity ratio of phycocyanin and its antioxidant activities. This will be further investigated for the development into anti-aging cosmetic products.
