❞ كتاب SV40 Protocols ❝  ⏤ Leda Raptis

❞ كتاب SV40 Protocols ❝ ⏤ Leda Raptis

نبذه 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. Leda Raptis - ❰ له مجموعة من الإنجازات والمؤلفات أبرزها ❞ SV40 Protocols ❝ ❱
من Biology Books علم الأحياء - مكتبة الكتب العلمية.

نبذة عن الكتاب:
SV40 Protocols

2001م - 1445هـ
نبذه 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.
.
المزيد..

تعليقات القرّاء:

Biologically

Biology is a natural science that is concerned with the study of life, its various forms and its function, how these organisms interact with each other and with the surrounding environment. The word biology in Greek is made up of two words: bio (βίος) meaning life. And loggia (-λογία) means science or study. Biology: the similarity of vegetation and animal cover on the edges of the African and American states, and the existence of the same fossil.


Branches of biology
Biology is an ancient science thousands of years old and modern biology began in the nineteenth century. This science has multiple branches. Among them are:

Anatomy
Botany
Biochemia
Biogeography
Biofisia
Cytology or cell science
Ecology or environmental science
 

 

نبذه 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.

Biology
Human biology
Who is the founder of biology?
The importance of biology
Areas of work in the field of biology
Theories of biology
Research on biology for the first grade of secondary school
Human biology



سنة النشر : 2001م / 1422هـ .
حجم الكتاب عند التحميل : 3.307 .
نوع الكتاب : pdf.
عداد القراءة: عدد قراءة SV40 Protocols

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Leda Raptis -

كتب Leda Raptis ❰ له مجموعة من الإنجازات والمؤلفات أبرزها ❞ SV40 Protocols ❝ ❱. المزيد..

كتب Leda Raptis