Roth AE86.1 User guide

Gel extraction kit Ultrafree® DA

DNA Extraction from Agarose Gels, Range: 100 –10.000 bp DNA

AE86.1

Introduction

This centrifugal filter device is designed to extract 100 to 10,000 bp DNA from agarose gel slices in one 10 minute
spin. DNA prepared with this device requires no further purification for use in cloning and radioisotopic or fluorescent
DNA sequencing. Due to the high resolving power of agarose gel electrophoresis, the small and large non-specific
amplification products that frequently interfere with cloning and sequencing after PCR* are completely removed from
your product.

* PCR (polymerase chain reaction) is covered by U.S. and other equivalent patents issued to Hoffman-La Roche, Inc.

Contents of Package Quantity/pk (preassembled)
Gel Nebulizer, Ultrafree-MC (0.45 µm Durapore) 50

and Microcentrifuge filtrate vials 50

Additional Materials Required

 TAE electrophoresis buffer (40 mM Tris-acetate, pH 8.0, 0.1 mM Na

2

EDTA).

Note: In principle, the Ultrafree-DA gel extraction kit works with all standard buffer systems like TBE or TAE buffers.
However, since gel extractions are generally performed prior to enzymatic processes done with the eluted DNA, we
recommend modified TAE (Roth TAElight, Art. No. 0122.1) rather than TBE for the following reasons:
(1) TBE buffer strongly inhibits DNA sequencing reactions while modified TAE buffer does not. (2) Modified TAE has
reduced contents of Na2-EDTA while regular TAE has 1.0 mM Na2-EDTA. This EDTA level will not interfere with the
magnesium concentration in sequencing reactions and other downstream enzymatic treatments, many that are dependent
on magnesium.

 Agarose for genetic engeneering (e.g. Carl Roth Agarose GTQ, Art. No. 6352.2)

Note: Low melting point agarose is not compatible with this protocol.

Also suitable for isolation of
DNA from polyacrylamide gels.
Use same protocol.

Instructions for use

Procedure to Use Ultrafree-DA

1. Electrophorese 30 µl of PCR product or other DNA

through a < 1.25% ordinary agarose gel, prepared in
TAE buffer with ethidium bromide (0.5 µg/mL).

2. Locate band of interest with the long-wavelength
ultraviolet lamp or transilluminator. With a razor blade,
cut out the slice of agarose (<100 µl at 100 mg)
containing the band of interest. The Ultrafree-DA is pre­assembled as follows:

3. Place gel slice into Gel Nebulizer and seal device with
the cap attached to vial.

4. Spin at 5,000 x g for 10 minutes. Centrifugation forces the agarose through the Gel Nebulizer, converting it to a
fine slurry that is captured by Ultrafree-MC. Extruded DNA in electrophoresis buffer passes through the
microporous membrane in Ultrafree-MC and collects in the filtrate vial.

5. Discard the Ultrafree-MC and Gel Nebulizer. You can sequence or clone the DNA in the filtrate without
further purification.

Technical Assistance

For more information, contact Carl Roth Gmbh + Co. KG (Karlsruhe, Germany, 0049 – 721 5606-0) or the Millipore
office nearest you.
In the U.S., call 1-800-MILLIPORE (1-800-645-5476). Outside the U.S., see your Millipore laboratory catalogue for the
phone number of the office nearest you. You can reach us by e-mail at [email protected] or visit our web
site (www.millipore.com). Millipore Corporation is pleased to provide internet access to Material Safety Data Sheets
(MSDS) for its products that contain hazardous materials.
To obtain any MSDS documents that may be associated with this product, go to the MSDS page of the Carl Roth
website (www.carlroth.com) or the Millipore website (www.millipore.com/msds.nsf/home).

Standard Warranty

Millipore Corporation ("Millipore") warrants its products will meet their applicable published specifications when used in accordance with their
applicable instructions for a period of one year from shipment of the products. MILLIPORE MAKES NO OTHER WARRANTY, EXPRESSED OR
IMPLIED. THERE IS NO WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE. The warranty provided herein and

the data, specifications and descriptions of Millipore products appearing in Millipore's published catalogues and product literature may not be altered
except by express written agreement signed by an officer of Millipore. Representations, oral or written, which are inconsistent with this warranty or such
publications are not authorized and if given, should not be relied upon. In the event of a breach of the foregoing warranty, Millipore's sole obligation shall
be to repair or replace, at its option, the applicable product or part thereof, provided the customer notifies Millipore promptly of any such breach. If after
exercising reasonable efforts, Millipore is unable to repair or replace the product or part, then Millipore shall refund to the customer all monies paid for
such applicable product or part. MILLIPORE SHALL NOT BE LIABLE FOR CONSEQUENTIAL, INCIDENTAL, SPECIAL OR ANY OTHER INDIRECT

DAMAGES RESULTING FROM ECONOMIC LOSS OR PROPERTY DAMAGE SUSTAINED BY ANY CUSTOMER FROM THE USE OF ITS
PRODUCTS. Millipore is a registered trademark of Millipore Corp. or an affiliated company. Ultrafree is a registered trademark of Millipore Corp. or an affiliated company.

SeaKem is a registered trademark of FMC BioProducts. 99467 Rev.C, 01/01

Gel extraction kit Ultrafree® DA 50 units AE86.1

Carl Roth GmbH + Co. KG

Schoemperlenstraße 3-5 • 76185 Karlsruhe
P.O. Box 100121 • 76231 Karlsruhe

Phone: [phone]
Fax: [phone]
[email protected] • www.carlroth.com sse 06/2021

Die Firma ist eine Kommanditgesellschaft mit Sitz in Karlsruhe, Reg. Gericht
Mannheim HRA 100055. Persönlich haftende Gesellschafterin ist die Roth Chemie
GmbH mit Sitz in Karlsruhe, Reg. Gericht Mannheim HRB 100428.
Geschäftsführer: André Houdelet