85034-538-74
Technical data and operating instructions
Vivacon® 500
For in vitro use only
2 |
Vivacon® 500 µl – Introduction Operation
Storage conditions | shelf life
Vivacon® ultrafiltration spin columns
should be stored at room temperature.
The devices should be used before the
expiry date printed on the box.
Introduction
Vivacon
®
500 concentrators are disposable
ultrafiltration devices optimally suited for
DNA and protein concentration. For optimal
performance with DNA and protein samples,
they are equipped with the patented regenerated cellulose membrane Hydrosart
®
.
Vivacon
®
500 can be used in a benchtop
fixed angle rotor, accepting 1.5 | 2.2 ml
centrifuge tubes.
Equipment required for
Vivacon
®
500
Centrifuge
Rotor type Fixed angle
Minimum 40°
rotor angle
Rotor cavity To fit 1.5 | 2.2 ml/11
mm
conical bottom tubes
1. When working with DNA samples, select
a molecular weight cut off (MWCO) which
retains the fragment size of double
stranded DNA (ds DNA) as shown in
Table 5. When working with proteins,
select a MWCO at least 50% smaller than
the molecular size of the protein of
interest.
2. Fill concentrator with up to maximum
volumes shown in Table 1. (Ensure lid
is fully sealed).
3. Insert assembled concentrator into
centrifuge.
4. Centrifuge at speeds recommended in
Table 2, taking care not to exceed the
maximum g force indicated by the MWCO.
5. Once the desired concentration is
achieved, (see Table 3 or 4 for guide to
concentration times), remove assembly
and recover sample by reverse spinning
the concentrate into a fresh collection
tube. In this procedure remove filtrate
tube, invert the concentrator body into
new filtrate tube and then spin at up
to 2,500 g for 2 minutes (or pulse for
20–30 seconds).
The filtrate can be sealed for storage by
closing the filtrate tube cap.
Note:
It is not possible to do a backflush
with the device.
Table 1: Technical Specifications
Concentrator capacity
Fixed angle rotor 0.5 ml
Dimensions
Total length 45 mm
(Concentration)
Total length (Back-spin) 47.5 mm
Width 12.4 mm
Active membrane area 0.32 cm
2
Hold-up volume < 5 µl
Dead stop volume 5 µl (40° rotor)
Materials of construction
Body Polycarbonate
Filtrate vessel Polypropylene
Membrane
2kDa - 100 kDa Hydrosart
®
125 kDa Cellulose
Acetate (CA)
O-ring Medical grade
silicone
Table 2: Recommended Spin Speed in
Fixed Angle Rotor (x g)
Membrane cut off For DNA For proteins
2 kDa MWCO 7,500 14,000
10 kDa MWCO 7,500 14,000
30 kDa MWCO 5,000 14,000
50 kDa MWCO 5,000 14,000
100 kDa MWCO 3,000 8,000
125 kDa MWCO 2,500* 5,000
Technical Specifications
Desalting | Buffer Exchange
1. Concentrate sample to desired level.
2. Empty filtrate container.
3. Refill concentrator with an appropriate
solvent.
4. Concentrate the sample again and repeat
the process until the concentration of
contaminating microsolute is sufficiently
reduced. Typically 3X wash cycles will
remove 99% of initial salt concentration.
Vivacon
®
500 Reverse Spinning
| 3
Hydrosart
membrane
Reverse
spinning
concentrate
into fresh
collection
tube
Collection tube
fits standard
1.5 | 2.2 ml tube
carriers
Filtrate can
be sealed
for storage
* Spin speed 2,500 + g for DNA samples > 900 bp when using a 125 kDa MWCO.
For DNA samples >650 bp, spin at 1,000 + g.
4 |
Usage Tips
1. Flow Rate
Filtration rate is affected by several parameters, including MWCO, porosity, sample concentration, viscosity, centrifugal force and
temperature. Expect significantly longer spin
times for starting solutions with over 5% solids. When operating at 4°C, flow rates are
approximately 1.5 times slower than at 25°C.
Viscous solutions such as 50% glycerine will
take up to 5 times longer to concentrate
than samples in a predominantly buffer
solution.
2. Pre-rinsing
Membranes fitted to Vivacon
®
concentrators
contain trace amounts of glycerine. Should
these interfere with analysis, they can be
removed by rinsing fill volume of buffer
solution or deionised water through the concentrator. Decant filtrate and concentrate
before processing sample solution. If you do
not want to use the pre-rinsed device immediately, store it in the refrigerator with buffer
or water covering the membrane surface.
Please do not allow the membrane to dry
out.
3. Sterilisation of Vivacon
®
Devices
Vivacon
®
devices should not be autoclaved
as high temperatures will substantially
increase membrane MWCO. To sterilise,
use a 70% ethanol solution or sterilising
gas mixture.
4. Chemical Compatibility
Vivacon
®
concentrators are designed for use
with biological fluids and aqueous solutions.
For chemical compatibility details, please
refer to Table 6.
5. Retention and Recovery
The membranes used in Vivacon
®
are
characterized by a molecular weight cut
off (MWCO). For proteins, it corresponds
to their ability to retain 90% of a molecule
with this nominal molecular weight. For
achieving better recovery, use a MWCO
which is 1 to 2 of the species weight you
need to concentrate.
For nucleic acid applications, strand length
is the most useful parameter for selecting
the Vivacon
®
device appropriate for a specific
application. However, other parameters
including DNA concentration, the magnitude
of the driving force (g-force) and the salt
concentration all act in concert to affect
DNA recovery. For characteristic recoveries
and concentration times, see Table 3 and 4.
For the correlation between MWCO and
nucleotide cut-off (bp), see Table 5.
Performance Characteristics
Table 3: Performance Characteristics Vivacon® 500 for DNA
Start volume 0.5 ml, sample concentration 50 ng/ml
Sample size (bp)
Time to concentrate up Concentrate g-force (xg)
to 30x [min.] at 20°C recovery %
2,000 MWCO 10 60 min 93% 7,500
10,000 MWCO 30 25 min 94% 7,500
30,000 MWCO 50 18 min 88% 5,000
50,000 MWCO 300 18 min 91% 5,000
100,000 MWCO 600 10 min 87% 3,000
125,000 MWCO 650 12 min 85% 2,000
125,000 MWCO 900 9 min 94% 3,000
Table 4: Performance Characteristics Vivacon
®
500 for Proteins
Start volume 0.5 ml, sample and concentration of proteins as specified in table
Sample
Time to concentrate up Concentrate g-force (xg)
to 30x [min.] at 20°C recovery %
2,000 MWCO 0.25 mg/ml 30 min 95% 14,000
cytochrome c
10,000 MWCO 0.25 mg/ml 15 min 92% 14,000
cytochrome c
30,000 MWCO 1.0 mg/ml BSA 10 min 95% 14,000
50,000 MWCO 1.0 mg/ml BSA 10 min 92% 14,000
100,000 MWCO 1.0 mg/ml 11 min 90% 8,000
bovine IgG
125,000 MWCO 1,0 mg/ml 10 min 81% 8,000
bovine IgG
Table 5: Conversion Table for Hydrosart
®
MWCO to Nucleotide Cut-off
Membrane MWCO Double-Stranded
Nucleotide Cut-off (bp)
Hydrosart
®
2 kDa > 10
Hydrosart
®
10 kDa > 30
Hydrosart
®
30 kDa > 50
Hydrosart
®
50 kDa > 300
Hydrosart
®
100 kDa > 600
Cellulose Acetate 125 kDa >650
| 5
6 |
Table 6: Chemical Compatibility (2hr contact time)
Hydrosart
®
Cellulose Acetate
Compatible pH range pH 1-9 pH 4-8
Acetic Acid (25.0%) OK NO
Acetone (10.0%) NO NO
Acetonitrile (10.0%) NO NO
Ammonium Hydroxide (5.0%) OK OK
Benzene (100%) NO NO
Chloroform (1%) OK OK
Dimethyl Formamide (10.0%) NO NO
Dimethyl Sulfoxide (5.0%) NO NO
EDTA (1.0 M) Yes Yes
Ethanol (70.0%) OK OK
Ethyl Acetate (100%) NO NO
Formaldehyde (30%) OK OK
Formic Acid (5.0%) OK ?
Glycerine (70%) OK OK
Guanidine HCI (6 M) OK ?
Hydrocarbons, aromatic NO NO
Hydrocarbons, chlorinated NO NO
Hydrochloric Acid (1 M) OK NO
Isopropanol (70%) OK OK
Lactic Acid (5.0%) OK NO
Mercaptoethanol (1.0 M) OK NO
Methanol (60%) OK OK
Nitric Acid (10.0%) NO NO
Phenol (1%) OK OK
Phosphate Buffer (1.0 M) OK OK
Sodium Chloride (5 M) Yes Yes
Sodium Dodecylsulfate (0.1 M) OK OK
Sodium Hydroxide (1.0 M) NO NO
Sodium Hypochlorite (200 ppm) NO NO
Sodium Nitrate (1.0%) OK ?
Tetrahydrofuran (5.0%) NO NO
Toluene (1.0%) NO NO
Trifluoroacetic Acid (10%) OK NO
TRIS Buffer, pH 7.2 - 9 (1.0 M) Yes Yes
Tween 20 (0.1%) OK OK
Triton X-100 (0.1%) OK OK
Urea (8 M) OK ?
OK = Acceptable ? = Questionable NO = Not recommended
FAQ
– DNA recovery is lower than expected
If the DNA sample contains a high salt concentration, dilute the sample.
Run the device at the recommended g-force.
– Can proteins be concentrated with Vivacon
®
?
Proteins can be concentrated with Vivacon
®
, using the guidelines on page 5 to choose the
correct MWCO. However, we recommend Vivaspin
®
500 for protein concentration due to faster
concentration achieved with a vertical membrane design for protein applications.
–
Sample runs to dryness.
Spinning for much longer than the recommended spin times can allow samples to go
to dryness. To recover the sample, add 10 µl of water or buffer to the device, vortex gently
for up to1 min and then recovery as normal.
– DNA recovery too low.
Spinning the sample at 1000 + g may result in higher DNA recoveries, when working close
to the membrane cut off limits, e.g. with a 650 bp DNA sample with a 125 kDa membrane.
–
No DNA signal visible after PCR reaction.
Using membrane cut offs smaller than 125 kDa can in some cases lead to concentration
of PCR inhibitors along with the sample DNA. Use the 125 kDa membrane cut off and spin
your samples at 2000 + g for optimal DNA recovery and sequencing results.
Additionally, 1-2 washes with buffer may be needed to remove the inhibitors.
Ordering Information
Vivacon
®
500 Qty. per box Prod. No.
2,000 MWCO 25 VN01H91
2,000 MWCO 100 VN01H92
10,000 MWCO 25 VN01H01
10,000 MWCO 100 VN01H02
30,000 MWCO 25 VN01H21
30,000 MWCO 100 VN01H22
50,000 MWCO 25 VN01H31
50,000 MWCO 100 VN01H32
100,000 MWCO 25 VN01H41
100,000 MWCO 100 VN01H42
125,000 MWCO 25 VN01H81
125,000 MWCO 100 VN01H82
125,000 MWCO 500 VN01H83
Vivacon
®
500 Qty. per box Prod. No.
Sample Kit L (4 units each of 2, 10, 30 K) 12 VN01HL12
Sample Kit H (4 units each of 30, 50, 100 K) 12 VN01HH12
| 7
Sartorius Stedim Lab Ltd
Sperry Way
Stonehouse Park
Gloucestershire
GL10 3UT, UK
www.sartorius.com
Copyright by
Sartorius Lab Instruments
GmbH & Co. KG, Goettingen,
Germany.
All rights reserved. No part
of this publication may
be reprinted or translated in
any form or by any means
without the prior written
permission of Sartorius Lab
Instruments GmbH & Co. KG.
The status of the information,
specifications and illustrations
in this manual is indicated
by the date given below.
Sartorius Lab Instruments
GmbH & Co. KG reserves the
right to make changes to the
technology, features,
specifications and design of the
equipment without notice.
Status:
June 2016,
Sartorius Lab Instruments
GmbH & Co. KG,
Goettingen, Germany
Printed in the EU on paper bleached
without chlorine.
Vivacon · W
Publication No.: SLL6002-e160607
Ver. 06 | 2016
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