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PCR试验的实验室参考手册
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PCR实验的实验室参考手册，PDF格式，可供实验室参考。 部分目录： PREFACE.................................................................................... 4 LAB SUPPLY SELECTION ........................................................ 5 LABORATORY GLOVES ................................................................ 5 PCR MICROTUBES ..................................................................... 9 PIPETTS & TIPS ........................................................................ 12 PCR SEALING & STORAGE PRODUCTS...................................... 16 PRIMERS .................................................................................. 19 PRIMER DESIGN TIPS ................................................................ 19 REVIEWS OF ON-LINE AND FREEWARE PRIMER DESIGN TOOLS...... 22 PCR METHODS ........................................................................ 32 STANDARD PCR ....................................................................... 32 LA PCR................................................................................... 33 HOT START PCR ...................................................................... 34 TOUCHDOWN PCR.................................................................... 39 RT PCR................................................................................... 40 IN SITU PCR............................................................................. 45 QUANTITATIVE PCR.................................................................. 49 PCR VARIABLES ..................................................................... 51 Primers................................................................................. 52 Polymerases ........................................................................ 52 Templates ............................................................................ 53 Magnesium ions................................................................... 54 Preincubation Temperatures and Times.............................. 55 Melting Temperatures and Times........................................ 55 Temperature - Annealing/Hybridization ............................... 56 Temperature - Extension/Polymerization............................. 56 Estimating extension times for LA PCR............................... 57 Cycles .................................................................................. 57 Reaction Volumes................................................................ 58 PCR POLYMERASES............................................................... 60 TAQ (THERMUS AQUATICUS)...................................................... 62 KLENTAQ (THERMUS AQUATICUS, N-TERMINAL DELETION MUTANT) ..63 STOFFEL FRAGMENT ................................................................. 64 TTH (THERMUS THERMOPHILUS).................................................64 PFU (PYROCOCCUS FURIOSUS) ..................................................65 VENT (THERMOCOCCUS LITORALIS)............................................66 DEEP VENT (PYROCOCCUS SPECIES GB-D) ...............................67 ULTMA (THERMOTOGA MARITIMA)..............................................68 PCR TROUBLESHOOTING......................................................69 Non-Specific Product Yields.................................................69 Little or No Product Yield......................................................70 Multiple Product Yields or High Molecular-Weight Smear is Observed..............................................................................72 Primer-Dimers ......................................................................73 About Inhibitors.....................................................................74 About Enhancers..................................................................74 How to Reduce Contamination.............................................76 Techniques Used to Prevent Contamination........................77 PCR INHIBITORS & ENHANCERS ..........................................79 GENERAL/INTRO, FECES, FORMALIN-FIXED PARAFFIN SAMPLE ...79 FECES, NORWALK VIRUS, ROTAVIRUS, URINE ............................82 RESPIRATORY FLUID, BLOOD, SHELLFISH...................................86 BODY FLUIDS AND FOOD............................................................90 PLANTS & SOIL .........................................................................94 SPECIAL PCR TOPICS.............................................................97 WHY AREN'T PCR-BASED DIAGNOSTICS MAIN STREAM?............97 PCR-BASED DETECTION KITS PART I.......................................100 PCR-BASED DETECTION KITS PART II.....................................103 FISH, PRINS & CYTOGENICS: DO YOU HAVE A LIGHT? ..........106 CLINICAL APPLICATIONS OF DOT BLOTS ...................................110 APPENDIX A ¨C UNITS AND FORMULAS..............................113 APPENDIX B ¨C THE GENETIC CODE....................................116 APPENDIX C ¨C ONLINE GENOMIC DATABASES................117 APPENDIX D ¨C PCR PRODUCT SUPPLIERS .......................124 APPENDIX E ¨C ONLINE BIOSUPPLIERS..............................137 APPENDIX F - CONTRIBUTORS ...........................................139 INDEX （温馨提示：资料在附件一） 渴望得到虫友们评价！ [ Last edited by judge0zz8 on 2008-1-22 at 10:23 ] |
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