A kind of ion exchange chromatography extraction method of abalone muscular dystrophy virus
A technology of ion exchange chromatography and abalone muscular dystrophy, which is applied in the field of ion exchange chromatography extraction of abalone muscular dystrophy virus, can solve the problems of limited sample processing capacity, inability to expand production, low virus purity, etc., and achieve convenient large-scale Sample preparation, shortened separation time, low cost effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0031] (1) Extraction of abalone muscular dystrophy virus
[0032] (1) Take 10g of virus-infected abalone tissues (all tissues except gonads), and grind them into powder under liquid nitrogen environment.
[0033] (2) Add 100mL SB buffer solution to the abalone tissue sample powder, and homogenize fully to obtain a homogenate. Among them, the pH of SB buffer is 7.5, and every 100ml contains 0.2M NaCl, 50mM Tris-HCl, 5mM CaCl 2 , 5 mM MgCl 2 .
[0034] (3) Centrifuge the homogenate twice with a common high-speed refrigerated centrifuge under the following conditions: centrifuge at 12,000 rpm for 30 min at 4°C, and collect the supernatant. The obtained supernatant was centrifuged at 10° C. at 40,000 rpm for 1 hour using an ultracentrifuge, and the supernatant was discarded. The obtained precipitate was the crude extract of abalone muscular dystrophy virus. The pellet was fully suspended with 10 ml of buffer A, which was 20 mM Tris, pH 8.0, to obtain a resuspension. The resu...
Embodiment 2
[0038] (1) Take 9g of virus-infected abalone tissues (all tissues except gonads), and grind them into powder under liquid nitrogen environment.
[0039](2) Add 100mL SB buffer solution to the abalone tissue sample powder, and homogenize fully to obtain a homogenate. Among them, the pH of SB buffer is 7.5, and every 100ml contains 0.15M NaCl, 45mM Tris-HCl, 4mM CaCl 2 , 4mM MgCl 2 .
[0040] (3) Centrifuge the homogenate twice with a common high-speed refrigerated centrifuge under the following conditions: centrifuge at 11,000 rpm for 25 minutes at 4°C, and collect the supernatant. The obtained supernatant was centrifuged at 10° C. at 30,000 rpm for 1.5 h in an ultracentrifuge, and the supernatant was discarded. The obtained precipitate was the crude extract of abalone muscular dystrophy virus. The pellet was fully suspended with 11 ml of buffer A, which was 25 mM Tris, pH 8.0, to obtain a resuspension. The resulting suspension was filtered with a 0.22um filter.
[0041] (...
Embodiment 3
[0044] (1) Take 10g of virus-infected abalone tissues (all tissues except gonads), and grind them into powder under liquid nitrogen environment.
[0045] (2) Add 100mL SB buffer solution to the abalone tissue sample powder, and homogenize fully to obtain a homogenate. Among them, the pH of SB buffer is 7.5, and every 100ml contains 0.3M NaCl, 55mM Tris-HCl, 6mM CaCl 2 , 6mM MgCl 2 .
[0046] (3) The homogenate was centrifuged twice in an ordinary high-speed refrigerated centrifuge under the following conditions: centrifuge at 13,000 rpm for 20 min at 4°C, and collect the supernatant. The obtained supernatant was centrifuged at 10° C. at 25,000 rpm for 2 hours in an ultracentrifuge, and the supernatant was discarded. The obtained precipitate was the crude extract of abalone muscular dystrophy virus. The pellet was fully suspended with 10 ml of buffer A, which was 20 mM Tris, pH 8.0, to obtain a resuspension. The resulting suspension was filtered with a 0.22um filter.
[00...
PUM
| Property | Measurement | Unit |
|---|---|---|
| diameter | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More