The viruses in the allantoic fluid were first inactivated using 0.05% BPL as described previously2 . To concentrate the viruses, allantoic fluids were clarified by centrifugation at 3441 × g at 4 °C for 30 min using a Sorvall Legend RT Plus Refrigerated Benchtop Centrifuge (Thermo Fisher Scientific). Clarified allantoic fluids were laid on top of a 20% sucrose cushion in NTE buffer (100 mM NaCl, 10 mM Tris-HCl, 1 mM EDTA, pH 7.4). Ultracentrifugation in a Beckman L7-65 ultracentrifuge at 112,499 × g for 2 hours at 4 °C using a Beckman SW28 rotor (Beckman Coulter) was performed to pellet the viruses through the sucrose cushion while soluble egg proteins were removed. The virus pellets were resuspended in PBS (pH 7.4). The total protein content was determined using the bicinchoninic acid assay (Thermo Fisher Scientific).
Sorvall legend rt plus refrigerated benchtop centrifuge
Manufactured by Thermo Fisher Scientific
The Sorvall Legend RT Plus Refrigerated Benchtop Centrifuge is a compact, high-performance laboratory centrifuge designed for a variety of applications. It features a refrigeration system to maintain sample integrity, and has a speed range of up to 15,000 rpm and a maximum relative centrifugal force of 21,382 x g.
Automatically generated - may contain errors
Lab products found in correlation
Beckman sw28 rotor, by Beckman Coulter (4 mentions)
Bicinchoninic acid bca assay, by Thermo Fisher Scientific (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
Bicinchoninic acid assay, by Thermo Fisher Scientific (2 mentions)
L7 65, by Beckman Coulter (1 mentions)
4 protocols using sorvall legend rt plus refrigerated benchtop centrifuge
1
Virus Concentration and Purification from Allantoic Fluid
2
Virus Purification from Allantoic Fluid
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Allantoic fluids were clarified by centrifugation at 4000 rpm using a Sorvall Legend RT Plus Refrigerated Benchtop Centrifuge (Thermo Fisher Scientific) at 4 °C for 30 min. Viruses in the allantoic fluid were pelleted through a 20% sucrose cushion in NTE buffer (100 mM NaCl, 10 mM Tris–HCl, 1 mM ethylenediaminetetraacetic acid (EDTA), pH 7.4) by centrifugation in a Beckman L7–65 ultracentrifuge at 25,000 rpm for 2 h at 4 °C using a Beckman SW28 rotor (Beckman Coulter, Brea, CA, USA). Supernatants were aspirated and the pellets were re-suspended in PBS (pH 7.4). The protein content was determined using the bicinchoninic acid (BCA) assay (Thermo Fisher Scientific).
3
Virus Concentration and Purification Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The viruses in the allantoic fluid were first inactivated using 0.05% beta-propiolactone (BPL) as described previously [23 ]. To concentrate the viruses, allantoic fluids were clarified by centrifugation at 4,000 rpm at 4°C for 30 min using a Sorvall Legend RT Plus Refrigerated Benchtop Centrifuge (Thermo Fisher Scientific). Clarified allantoic fluids were laid on top of a 20% sucrose cushion in PBS (Gibco). Ultracentrifugation in a Beckman L7–65 ultracentrifuge at 25,000 rpm for 2 hours at 4°C using a Beckman SW28 rotor (Beckman Coulter, CA, USA) was performed to pellet the viruses through the sucrose cushion while soluble egg proteins were removed. The virus pellets were re-suspended in PBS (pH 7.4). The total protein content was determined using the bicinchoninic acid (BCA) assay (Thermo Fisher Scientific).
4
Virus Purification from Allantoic Fluid
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The viruses in the allantoic fluid were first inactivated using 0.05% beta-propiolactone (BPL) as described previously (18 (link)). To concentrate the viruses, allantoic fluids were clarified by centrifugation at 4,000 rpm at 4°C for 30 min using a Sorvall Legend RT Plus Refrigerated Benchtop Centrifuge (Thermo Fisher Scientific). Clarified allantoic fluids were laid on top of a 20% sucrose cushion in PBS (Gibco). Ultracentrifugation in a Beckman L7-65 ultracentrifuge at 25,000 rpm for 2 h at 4°C using a Beckman SW28 rotor (Beckman Coulter, CA, USA) was performed to pellet the viruses through the sucrose cushion while soluble egg proteins were removed. The virus pellets were re-suspended in PBS (pH 7.4). The total protein content was determined using the bicinchoninic acid (BCA) assay (Thermo Fisher Scientific).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!