Q: What is the optimal harvest time for maximum Adenovirus yield?
In general, a much higher virus yield
is obtained using a 4-5 day procedure, and for this
reason alone we recommend it. In general, a 48 hour procedure
results in a higher level of contaminating cellular
and viral DNA relative to the 4-5 day procedure.
Q: How much Adenovirus will the Adenopure® kit
purify?
The Adenopure® purification procedure
will yield between 1x1012 and 5x1012 viral particles. Of course this is dependant upon
loading at least that much virus onto the Virus-Binding
Module of the Adenopump® Purification Unit.
Each kit contains enough reagents for (3) purification
procedures.
Q: Can I pass more than (5)
150cm2 plates worth of infected cell extract
through an Adenopump® virus binding module?
The Adenopure® purification procedure
has been optimized to process (5) 150cm2 plate equivalents of infected cell extract. If more
plate equivalents of infected cell extract are used
in a purification procedure, the contaminating free
cellular and viral DNA may not be totally degraded
and the amount and purity the resulting Adenovirus
preparation may be compromised. In addition, the
user will run short on some of the buffers supplied
in the kit.
Q: How critical is it to
add the right amount of 10X Dilution Buffer?
It is extremely critical to add the
right amount of 10X Dilution Buffer to the sample
as this buffer allows efficient binding of Adenovirus
to the Virus-Binding Module. The addition of an incorrect
amount of 10X Dilution Buffer will lead to a decrease
in the strength of interaction of Adenovirus with
the Virus-Binding Module as well as allowing a different
spectrum of contaminating proteins to compete with
the Adenovirus for binding to the Virus-Binding Module.
Q: Can I reuse an Adenopump
Purification Unit?
The Adenopump Purification Units
are intended for single use only and should be disposed
of after a purification procedure in an appropriate
biohazardous waste receptacle.
Q: What happens if I push
the sample through the Adenopump Purification
Unit at a higher flow rate than 10ml/minute?
The efficiency of binding of Adenovirus
to the Virus-Binding Module will decrease with increasing
flow rate, and the overall recovery of purified Adenovirus
will decrease as a result.
Q: What is the composition
of the buffers in the kit?
The formulation of the Adenopure Kit buffers contained
within the Adenopure kit is proprietary. MSDS are available for review.
For more information please contact (610) 640-0800 or email cs@puresyn.com