PEG Virus Precipitation Kit
Catalog#: K904-200 | Size: 200 preparations
PEG病毒濃度操作方法
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Virus Precipitation Protocol:
The following protocol is designed for 10 ml virus
solution. You can proportionally adjust the
volumes according to your sample volume. The kit is also
available for larger volume
samples. Please inquire.
1. Infect cells or transfect and allow maximum virus
accumulation.
2. For mammalian cell - virus or insect - baculovirus,
centrifuge culture at 3,200 x g for 15 min
at 4°C
to remove cells debris. For bacterial phage, centrifuge at 16,000 x g for 15
min at 4°
C to remove cells debris.
3. Collect supernatant and add 2.5 ml of PEG solution A
to 10 ml of virus supernatant.
Refrigerate overnight (stable up to 2 days at 4°C ).
4. Centrifuge at 3,200 x g for 30 min at 4°C , carefully remove supernatant by
aspiration. The
beige or white pellet is the virus.
5. Suspend the virus pellet in 100 μl Virus
Resuspension Solution. Aliquot the virus and store
at -70°C for future use.
病毒濃縮後如何移除溶液中的PEG
i) Add 1 volume of solution containing 4 M KCl and 50 mM Tris-HCl, pH7.2 (not
provided) to 3 volumes of the concentrated virus
suspension.
ii) Alternatively, add solid KCl into the virus
suspension to a final concentration of
1 M.
iii) Let stand on ice for 15 - 30 min.
iv) Spin at 12,000 x g for 10 min at 4°C to remove the precipitate.
v) Carefully collect the virus
supernatant. Aliquot and store at -70°C
for future use.
文獻References:
1. Lech K. Current Protocols in Molecular biology
(1990) 1.13.1 -10
2. Kimlton C.P,
Corbitt G, Morris D.J. Journal of Virological Merthods. (1990) 28: 141-
146.
3. Colombet J.
Robin A, Lavie L, et al. Journal of Virological Merthods. (2007) 71: 212-
219.
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