端粒酶检测方法-s7700 trapeze telomerase detection kit.docVIP

TRAPEZE? Telomerase Detection Kit S7700 FOR RESEARCH USE ONLY Not for use in diagnostic procedures USA Canada Phone: +1(800) 437-7500 ? Fax: +1 (951) 676-9209 ? Europe +44 (0) 23 8026 2233 Australia +61 3 9839 2000 ? Germany +49-6192-207300 ? ISO Registered Worldwide ? custserv@ ? techserv@) This page left blank intentionally. _______________________________________________________________ TABLE OF CONTENTS I. INTRODUCTION 1 Using this Manual 1 Background 1 Principles of the Technique 2 Fig. 1A: TRAPEZE? Gel-Based Telomerase Detection Kit Assay Scheme 3 Fig. 1B: TRAPEZE? Gel-Based Telomerase Detection Kit Assay Gel 3 II. KIT COMPONENTS 4 Precautions 5 Materials Required But Not Supplied 5 III. TRAPEZE? GEL-BASED TELOMERASE DETECTION KIT PROCEDURE 7 Fig. 2: TRAPEZE? Gel-Based Telomerase Detection Kit Flow Chart 7 Extract Preparation 8 Experimental Design 9 Controls 9 Assay Design 11 Detection Methods 11 TRAPEZE? Gel-Based Telomerase Detection Kit Assay 13 Radioisotopic Detection 13 Non-radioactive Detection by Staining 15 IV. DATA ANALYSIS 17 Visual Analysis 17 Quantitation 18 Estimation of Processivity 19 Fig. 3A: Quantitation Curve 19 Fig. 3B: Quantitation of Telomerase Activity 20 Sensitivity and Specificity of the Assay 20 i V. TROUBLESHOOTING 21 Fig. 4: Troubleshooting Examples 25 VI. APPENDIX 27 Laboratory Setup and Precautions 27 Fig. 5: TRAPEZE? Gel-Based Telomerase Detection Kit Station Setup 28 TRAPEZE? Gel-Based Telomerase Detection Kit Station Setup (Area 1) 29 Determination of Protein Concentration 29 Checking the Efficiency of End-Labeling 30 Removing PCR Inhibitors 31 Enhancing Detection Sensitivity 32 Reagent and Buffer Preparation 33 VII. REFERENCES 35 References Citing the TRAPEZE? Gel-Based Telomerase Detection Kit 35 References Cited in the Manual 36 Disclaimers 39 Warranty 39 ii I. INTRODUCTION Using This Manual This manual accommodates both the novice and the experienced TRAPEZE? Kit user. These protocols