PCR实验的实验室参考手册，PDF格式，可供实验室参考。

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 Contamination76
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, SHELLFISH86
BODY FLUIDS AND FOOD90
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 FORMULAS113
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 BIOSUPPLIERS137
APPENDIX F - CONTRIBUTORS ...139
INDEX