📘 ❞ SV40 Protocols ❝ كتاب ــ Leda Raptis اصدار 2001
Biology Books - 📖 ❞ كتاب SV40 Protocols ❝ ــ Leda Raptis 📖
█ _ Leda Raptis 2001 حصريا كتاب SV40 Protocols 2024 Protocols: نبذه عن الكتاب: 3 2 Virus Stock Production 1 Prepare 80% to 100% confluent T75 flasks of BS C 1 or CV cells (see Note 1) 2 Remove medium from dishes and replace with mL MEM 2% FBS containing the appropriate dilution plaque forming units (PFU’s) achieve a multiplicity infection (MOI) between 0 01 2) 3 Allow proceed for h in an incubator while rocking plates at 15 min intervals assure complete coverage even distribution over monolayer 4 After this period, should be added each plate obtain a final volume 10 per cm 5 4 d incubation, replaced fresh 2% FBS point, checked daily signs cytopathic effects (CPE) by comparing infected monolayers uninfected control (see Note 3 Fig 6 When CPE develops point destruction with floating clumps cells, place –20°C overnight until completely frozen then thaw room temperature Repeat total three freeze thaw cycles 7 This is viral stock It will contain cellular debris, which can removed by centrifugation 200 RCF 5 min if desired Titering Plaque Assay 1 serial dilutions virus putting 20 solution into vortex vigorously (10–2 dilution) Repeat procedure using immediately preceding the stock make 10–4 10–6 10–7 10–8 made by using 4) Use infect 6 freshly of BS after removing old Be sure include dish with 1 without as mock Biology Books مجاناً PDF اونلاين Biologically Biology natural science that concerned study life, its various forms function, how these organisms interact other surrounding environment The word biology Greek up two words: bio (βίος) meaning life And loggia ( λογία) means Biology: similarity vegetation animal cover on edges African American states, existence same fossil Branches biology Biology ancient thousands years modern began nineteenth century has multiple branches Among them are: Anatomy Botany Biochemia Biogeography Biofisia Cytology cell science Ecology environmental science Development Embryology embryology Genetics genetics Histology histology Anthropology anthropology Microbiology bacteriology Molecular Biology Physiology functions organs Taxonemia taxonomy Virology virology Zoology zoology
كتاب
SV40 Protocols
ــ Leda Raptis
صدر 2001م
كتاب
SV40 Protocols
ــ Leda Raptis
صدر 2001منبذه عن الكتاب:
3.2. SV40 Virus Stock Production
1. Prepare 80% to 100% confluent T75 flasks of BS-C-1 or CV-1 cells (see Note 1).
2. Remove medium from dishes and replace with 2 mL of MEM-2% FBS containing the appropriate dilution of SV40 plaque forming units (PFU’s) to achieve a
multiplicity of infection (MOI) between 0.1 and 0.01 (see Note 2).
3. Allow infection to proceed for 2 h in an incubator while rocking plates at 15-min
intervals to assure complete coverage and even distribution over the monolayer.
4. After this period, MEM-2% FBS should be added to each plate to obtain a final
volume of 10 mL of medium per 10-cm plate.
5. After 4 d of incubation, the medium should be replaced with fresh MEM-2%
FBS. After this point, cells should be checked daily for signs of cytopathic
effects (CPE) by comparing infected monolayers to the uninfected control (see
Note 3 and Fig. 1).
6. When CPE develops to the point of complete destruction of the monolayer with
floating clumps of cells, place flasks at –20°C overnight or until completely frozen and then thaw at room temperature. Repeat for a total of three freeze/thaw
cycles.
7. This is the viral stock. It will contain cellular debris, which can be removed by
centrifugation at 200 RCF for 5 min if desired.
3.3. Titering SV40 by Plaque Assay
1. Prepare serial dilutions of the SV40 virus stock in MEM-2% FBS by putting
20 mL of stock solution into 2 mL of MEM-2 and vortex vigorously (10–2 dilution).
Repeat this procedure using the immediately preceding dilution in place of the
stock to make 10–4 and 10–6 dilutions. 10–7 and 10–8 dilutions should be made by
using 200 mL of the preceding dilutions in 2 mL of MEM-2% FBS (see Note 4).
2. Use 1 mL of each dilution to infect 6-cm dishes of freshly confluent dishes of
BS-C-1 or CV-1 after removing the old medium. Be sure to include a dish with
1 mL of MEM-2% FBS without virus as mock control.
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