Bio-Rad Fluorescent Assay Kits User Manual

Bio-Rad
Laboratories

CE dsDNA 1000

U.S. (800) 4BIORAD California [phone] Australia 02-9914-2800 Austria
(1) 877 89 01 Belgium 09-385 55 11 Canada [phone] China [phone]
Denmark 39 17 9947 Finland 90 804 2200 France (1) 49 60 68 34 Germany 089
31884-0 India 91-11-461-0103 Italy 02-21609 1 Japan 03-5811-6270 Hong Kong
7893300 The Netherlands 0318-540666 New Zealand 09-443 3099 Singapore (65)
272-9877 Spain (91) 661 70 85 Sweden 46 (0) 735 83 00 Switzerland 01-809 55 55
United Kingdom 0800 181134

Life Science
Group

SIG 020996 Printed in USA

Fluorescent Detection and

Analysis Kit

Instruction Manual

Catalog Number

148-4133

4006108 Rev B

For Technical Service Call Your Local Bio-Rad Office

or in the U.S. Call 1-800-4BIORAD (1-800-424-6723)

Table of Contents

Section 1 Introduction ..............................................................1

Section 2 CE dsDNA 1000 Fluorescent Detection and

Kit Components......................................................2

2.1 Capillary Preparation and Use ..............................................2

2.2 Using the CE dsDNA 100 Base Pair Ladder ..................3

2.3 Sample Preparation..........................................................4

2.4 Buffer Preparation for Resolving Fragments

of 100–1,000 Base Pairs........................................................4

Section 3 Analysis Conditions on the

BioFocus

3.1 Configuration......................................................................5

3.2 dsDNA Analysis Method ..................................................6

3.3 Analysis of Multiple Samples............................................7

Section 4 System Shutdown Method......................................8

Section 5 Optimization of Nucleic Acid Separations ........10

5.1 Shortening Analysis Time................................................10

5.2 Increasing Resolution ......................................................10

5.3 Regenerating a Plugged Capillary ..................................11

Section 6 Product Information............................................12

®

System ......................................................5

Section 1
Introduction

The CE dsDNA 1000 Fluorescent Detection and Analysis Kit
is optimized for use with the BioFocus®capillary electrophoresis
laser induced fluorescence LIF2detection system and resolves
double-stranded (ds) DNA fragments ranging in size from 100 to
1,000 base pairs. The kit is useful for a variety of applications
including determination of PCR*yields, optimizing PCR conditions,
and characterizing restriction enzyme digests. High performance
capillary electrophoresis is an ideal approach for nucleic acid
separations, providing rapid analysis, quantitative information,
minimal consumption of sample, and direct detection of resolved
fragments.

High resolution DNA separations are achieved by dynamic
sieving**capillary electrophoresis, a technique that incorporates a
hydrophilic polymer in the electrophoresis buffer . The polymer -con­taining buffer is replenished between each analysis, providing
reproducibility of migration time and peak area. The analysis
employs BioCap™capillaries which are coated internally with a
novel polymer (polyAAEE), insuring high resolution separations
over many runs.

SYBR®Green I, the fluorescent dye included in the kit, is an
intercalating monomeric dye with the excitation maximum close
to the 488 nm line of the argon ion laser, extremely low intrinsic
fluorescence, and very high affinity for dsDNA.

1