Agilent High Sensitivity DNA Kit Guide

Agilent High Sensitivity DNA Kit Guide

Agilent High Sensitivity DNA

Agilent Technologies

Notices

? Agilent Technologies, Inc. 2009, 2013 No part of this manual may be reproduced in any form or by any means (including electronic storage and retrieval or translation into a foreign language) without prior agreement and written consent from Agilent Technologies, Inc. as governed by United States and international copyright laws.

Manual Part Number

G2938-90321 Rev. B

Edition

11/2013 Printed in Germany Agilent Technologies Hewlett-Packard-Strasse 8 76337 Waldbronn

Resarch Use Only

Not for use in Diagnostic Procedures.

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Safety Notices

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Agilent High Sensitivity DNA

Contents

Contents

1 Agilent High Sensitivity DNA Kit 4

2 Equipment Required for a High Sensitivity DNA Assay 6

3 Setting up the Assay Equipment and Bioanalyzer 7 Setting up the Assay Equipment and Bioanalyzer 7 Setting up the Chip Priming Station 8 Setting up the Bioanalyzer 9 Vortex Mixer 9 Starting the 2100 Expert Software 10

4 Essential Measurement Practices 11

5 Agilent High Sensitivity DNA Assay Protocol 12 Preparing the Gel-Dye Mix 12 Loading the Gel-Dye Mix 14 Loading the Marker 15 Loading the Ladder and the Samples 16 Inserting a Chip in the Agilent 2100 Bioanalyzer 17 Starting the Chip Run 18 Cleaning Electrodes after a High Sensitivity DNA Chip Run 20

6 Checking Your Agilent High Sensitivity DNA Assay Results 21 High Sensitivity DNA Ladder Well Results 21 High Sensitivity DNA Sample Well Results 23

Agilent High Sensitivity DNA

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Agilent High Sensitivity DNA

Agilent High Sensitivity DNA Kit

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Table 1 Agilent High Sensitivity DNA Kit

Agilent High Sensitivity DNA Kit (reorder number 5067-4626)

DNA Chips

Agilent High Sensitivity DNA Reagents (reorder number 5067-4627)

10 High Sensitivity DNA Chips

(yellow) High Sensitivity DNA Ladder

1 Electrode Cleaner

(green) High Sensitivity DNA Markers 35/10380 bp (4 vials)

Syringe Kit

(blue) High Sensitivity DNA Dye Concentrate 1(1 vial)

1 Syringe

(red) High Sensitivity DNA Gel Matrix (2 vials)

2 Spin Filters

1 "This product is provided under a license by Life Technologies Corporation to Agilent Technologies. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only as described in accompanying product literature. The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell or otherwise transfer this product or its components to any third party, or use for any use other than use in the subfields of research and development, quality control, forensics, environmental analysis, biodefense or food safety testing. For information on purchasing a license to this product for purposes other than described above contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354"

Agilent Technologies

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Agilent High Sensitivity DNA Kit 1

Table 2 Physical Specifications

Type Analysis run time Number of samples Sample volume Kit stability

Specification 45 minutes 11 samples/chip 1 ?l 4 months (Storage temp. see individual box!)

Table 3 Analytical Specifications

Type

Agilent High Sensitivity DNA Assay

Sizing range

50-7000 bp

Typical sizing resolution

50-600 bp:? 10 % 600-7000 bp:? 20 %

Sizing accuracy

? 10 % CV (for ladder as sample)

Sizing reproducibility

5 % CV (for ladder as sample)

Quantitation accuracy

20 % CV (for ladder as sample)

Quant. reproducibility

50-2000 bp: 15 % CV; 2000-7000 bp: 10 % CV (for ladder as sample)

Quantitative range

5-500 pg/?l (for ladder as sample)

Maximum salt1

10 mM Tris and 1 mM EDTA

1 Due to the high sensitivity of the assay, different ions and higher salt concentrations might influence the performance of the assay.

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