Sartorius Vivacon 500 Bedienungsanleitung

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 regen-

erated 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

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 cm2

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 parame-

ters, including MWCO, porosity, sample con-

centration, viscosity, centrifugal force and

temperature. Expect significantly longer spin

times for starting solutions with over 5% sol-

ids. 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 con-

centrator. Decant filtrate and concentrate

before processing sample solution. If you do

not want to use the pre-rinsed device imme-

diately, 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

Sartorius Lab Instruments

GmbH & Co. KG, Goettingen,

Germany.

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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