GE 100 reactions 79770, 188403, 33, 50 reactions 79760, 50 reactions AH9539, 500 reactions, 79750 Figure

0.5pmol

1pmol

2pmol

8pmol

300 bases

150 bases

Figure 1. Excess template DNA can reduce sequence extension lengths. In cases where 2pmol or more template DNA are sequenced, the supply of nucleotides can be exhausted before extensions reach suitable length for optimal sequencing. These sequences were run using up to 16∝g (8pmol) of M13mp18 DNA template.

Contents

STORAGE CONTENTS COMPONENTS OF THE KIT QUALITY CONTROL SAFETY WARNINGS AND PRECAUTIONS INTRODUCTION Chain termination sequencing Page MATERIALS NOT SUPPLIED Necessary reagents: Necessary equipment: PROTOCOL 3.Cycling termination reactions SUPPLEMENTARY INFORMATION General guidelines Preparation of template DNA Preparation of single-strandedtemplate DNA Preparation of double-strandedplasmid DNA Cycle conditions and template quantity Cycling temperatures Number of cycles and quantity of template Designing a new sequencing primer Figure Sequencing PCR Products Enzymatic pre-treatmentof PCR products Elimination of compressions Page Reading farther from or closer to the primer Denaturing gel electrophoresis Gel electrophoresis reagents Buffers Gel recipes (for 100ml of gel solution) General guidelines for electrophoresis TROUBLESHOOTING Film blank or very faint Bands faint near the primer Bands smeared Bands appear across all 4 lanes CONTROL DNA SEQUENCE REFERENCES RELATED PRODUCTS Kits and Enzymes PCR Clean-up USB Ultrapure reagents for DNA sequencing USB CORPORATION Material safety data sheet HAZARDS IDENTIFICATION FIRST-AIDMEASURES FIRE-FIGHTINGINFORMATION ACCIDENTAL RELEASE MEASURES HANDLING AND STORAGE

GE manual Figure

Models: 100 reactions 79770 188403 33 50 reactions 79760 50 reactions AH9539 500 reactions 79750