Calculations for Molecular Biology and Biotechnology: A Guide to Mathematics in the Laboratory

**Calculations in Molecular Biology and Biotechnology: A Guide to Mathematics in the Laboratory is the first comprehensive guide devoted exclusively to calculations encountered in the genetic engineering laboratory. Mathematics, as a vital component of the successful design and interpretation of basic research, is used daily in laboratory work. This guide, written for students, technicians, and scientists, provides example calculations for the most frequently confronted problems encountered in gene discovery and analysis. The text and sample calculations are written in an easy-to-follow format. It is the perfect laboratory companion for anyone working in DNA manipulation and analysis. \*A comprehensive guide to calculations for a wide variety of problems encountered in the basic research laboratory. \* Example calculations are worked through from start to finish in easy-to-follow steps \* Key chapters devoted to calculations encountered when working with bacteria, phage, PCR, radioisotopes, recombinant DNA, centrifugation, oligonucleotides, protein, and forensic science. \*Written for students and laboratory technicians but a useful reference for the more experienced researcher. \*A valuable teaching resource.** Front Cover......Page 1 Calculations in Molecular Biology and Biotechnology......Page 4 Copyright Page......Page 5 Contents......Page 8 Foreword......Page 15 Significant Digits......Page 18 Rounding Off Significant Digits in Calculations......Page 19 Expressing Numbers in Scientific Notation......Page 21 Converting Numbers from Scientific Notation to Decimal Notation......Page 23 Adding and Subtracting Numbers Written in Scientific Notation......Page 25 Multiplying and Dividing Numbers Written in Scientific Notation......Page 26 Metric Prefixes......Page 30 Conversion Factors and Canceling Terms......Page 31 Calculating Dilutions: A General Approach......Page 35 Concentrations by a Factor of X......Page 37 Preparing Percent Solutions......Page 39 Diluting Percent Solutions......Page 40 Moles and Molecular Weight: Definitions......Page 44 Molarity......Page 45 Diluting Molar Solutions......Page 48 Converting Molarity to Percent......Page 49 Converting Percent to Molarity......Page 50 Normality......Page 51 PH......Page 52 pKa and the Henderson–Hasselbalch Equation......Page 56 The Bacterial Growth Curve......Page 59 Manipulating Cell Concentration......Page 63 Plotting OD550 vs. Time on a Linear Graph......Page 65 Plotting the Logarithm of OD550 vs. Time on a Linear Graph......Page 66 Plotting the Log of Cell Concentration vs. Time......Page 68 Calculating Generation Time......Page 69 Plotting Cell Growth Data on a Semilog Graph......Page 72 Determining Generation Time Directly from a Semilog Plot of Cell Concentration vs. Time......Page 76 Plotting Cell Density versus OD550 on a Semilog Graph......Page 77 The Fluctuation Test......Page 78 Fluctuation Test Example......Page 80 Variance......Page 81 Measuring Mutation Rate......Page 83 Measuring Cell Concentration on a Hemocytometer......Page 92 Multiplicity of Infection......Page 94 Probabilities and Multiplicity of Infection......Page 96 Measuring Phage Titer......Page 102 Diluting Bacteriophage......Page 103 Measuring Burst Size......Page 104 Quantitation of Nucleic Acids by Ultraviolet Spectroscopy......Page 107 Determining the Concentration of Double-Stranded DNA......Page 108 Using Absorbance and an Extinction Coefficient to Calculate Double-Stranded DNA Concentration......Page 111 Calculating DNA Concentration as a Millimolar (mM) Amount......Page 113 Determining the Concentration of Single-Stranded DNA Molecules......Page 114 Oligonucleotide Quantitation......Page 116 Measuring RNA Concentration......Page 120 Molecular Weight, Molarity, and Nucleic Acid Length......Page 121 Estimating DNA Concentration on an Ethidium Bromide–Stained Gel......Page 125 Using Radioactivity: The Curie......Page 126 Estimating Plasmid Copy Number......Page 127 Labeling DNA by Nick Translation......Page 129 Random Primer Labeling of DNA......Page 131 Labeling 3' Termini with Terminal Transferase......Page 136 cDNA Synthesis......Page 138 Homopolymeric Tailing......Page 145 In Vitro Transcription......Page 150 Synthesis Yield......Page 153 Overall Yield......Page 156 Stepwise Yield......Page 157 Calculating Micromoles of Nucleoside Added at Each Base Addition Step......Page 159 Template and Amplification......Page 160 Exponential Amplification......Page 162 PCR Efficiency......Page 164 Calculating the Tm of the Target Sequence......Page 168 Primers......Page 170 Primer Tm......Page 175 dNTPs......Page 182 DNA Polymerase......Page 185 Quantitative PCR......Page 188 Restriction Endonucleases......Page 203 The Frequency of Restriction Endonuclease Cut Sites......Page 205 Calculating the Amount of Fragment Ends......Page 206 Ligation......Page 209 Transformation Efficiency......Page 224 Genomic Libraries: How Many Clones Do You Need?......Page 225 cDNA Libraries: How Many Clones Are Enough?......Page 227 Expression Libraries......Page 228 Screening Recombinant Libraries by Hybridization to DNA Probes......Page 229 Sizing DNA Fragments by Gel Electrophoresis......Page 241 Generating Nested Deletions Using Nuclease BAL 31......Page 254 Protein Quantitation by Measuring Absorbance at 280 nm......Page 259 Using Absorbance Coefficients and Extinction Coefficients to Estimate Protein Concentration......Page 260 Relating Absorbance Coefficient to Molar Extinction Coefficient......Page 262 Determining a Protein's Extinction Coefficient......Page 263 Relating Concentration in Milligrams per Milliliter to Molarity......Page 265 Protein Quantitation Using A280 When Contaminating Nucleic Acids Are Present......Page 266 Protein Quantitation at 205 nm......Page 267 Protein Quantitation at 205 nm When Contaminating Nucleic Acids Are Present......Page 268 Measuring Protein Concentration by Colorimetric Assay- The Bradford Assay......Page 269 Using β-Galactosidase to Monitor Promoter Activity and Gene Expression......Page 271 Specific Activity......Page 274 The CAT Assay......Page 277 Use of Luciferase in a Reporter Assay......Page 282 In Vitro Translation–Determining Amino Acid Incorporation......Page 283 Relative Centrifugal Force (g Force)......Page 287 Converting g Force to Revolutions per Minute......Page 289 Determining g Force and Revolutions per Minute by Use of a Nomogram......Page 290 Calculating Sedimentation Times......Page 292 Alleles and Genotypes......Page 295 Calculating Genotype Frequencies......Page 297 Calculating Allele Frequencies......Page 298 The Hardy–Weinberg Equation and Calculating Expected Genotype Frequencies......Page 299 The Chi-Square Test: Comparing Observed to Expected Values......Page 303 Sample Variance......Page 307 Sample Standard Deviation......Page 308 Pi: The Power of Inclusion......Page 309 Pd: The Power of Discrimination......Page 310 DNA Typing and a Weighted Average......Page 311 The Multiplication Rule......Page 312 Index......Page 314 Calculations in Molecular Biology and Biotechnology: A Guide to Mathematics in the Laboratory is the first comprehensive guide devoted exclusively to calculations encountered in the genetic engineering laboratory. Mathematics, as a vital component of the successful design and interpretation of basic research, is used daily in laboratory work. This guide, written for students, technicians, and scientists, provides example calculations for the most frequently confronted problems encountered in gene discovery and analysis. The text and sample calculations are written in an easy-to-follow format. It is the perfect laboratory companion for anyone working in DNA manipulation and analysis.

*A comprehensive guide to calculations for a wide variety of problems encountered in the basic research laboratory.
* Example calculations are worked through from start to finish in easy-to-follow steps
* Key chapters devoted to calculations encountered when working with bacteria, phage, PCR, radioisotopes, recombinant DNA, centrifugation, oligonucleotides, protein, and forensic science.
*Written for students and laboratory technicians but a useful reference for the more experienced researcher.
*A valuable teaching resource. This book provides example calculations for the most commonly encountered problems in gene discovery, analysis, and other areas of biotechnology. In addition to showing how to perform key calculations, it emphasizes mastery of basic theoretical and laboratory principles