| Preface |
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vii | |
| Contributors |
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xiii | |
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PART I TECHNOLOGY PREPARATION |
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1 Preparation of Single-Stranded DNA for Pyrosequencing by LATE-PCR |
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3 | (10) |
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2 A Simplified Protocol for Preparing Pyrosequencing Templates Based on LATE-PCR Using Whole Blood as Starting Material Directly |
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13 | (10) |
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3 Improvement of LATE-PCR to Prepare Pyrosequencing Template |
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23 | (8) |
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4 Pyrosequencing Templates Generated by Nicking PCR Products with Nicking Endonuclease |
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31 | (10) |
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5 Pyrosequencing Templates Generated by Asymmetric Nucleic Acid Sequence-Based Amplification (Asymmetric-NASBA) |
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41 | (10) |
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6 Gel Immobilization of Acrylamide-Modified Single-Stranded DNA Template for Pyrosequencing |
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51 | (16) |
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7 Multiplex PCR Based on a Universal Biotinylated Primer to Generate Templates for Pyrosequencing |
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67 | (12) |
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PART II TEMPLATE INNOVATION |
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8 A Novel Pyrosequencing Principle Based on AMP-PPDK Reaction for Improving the Detection Limit |
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79 | (16) |
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9 Pyrosequencing Chemistry Coupled with Modified Primer Extension Reactions for Quantitative Detection of Allele Frequency in a Pooled Sample |
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95 | (20) |
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10 A Gel-Free SNP Genotyping Method Based on Pyrosequencing Chemistry Coupled with Modified Primer Extension Reactions Directly from Double-Stranded PCR Products |
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115 | (14) |
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11 Improvement of Pyrosequencing to Allow Multiplex SNP Typing in a Pyrogram |
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129 | (16) |
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12 Improvement of Pyrosequencing Sensitivity by Capturing Free Adenosine 5'-Phosphosulfate with Adenosine Triphosphate Sulfurylase |
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145 | (10) |
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13 Using Polymerase Preference Index to Design imLATE-PCR Primers for an Efficient Pyrosequencing |
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155 | (14) |
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PART III REAGENTS PREPARATION |
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14 Preparation of Thermal-Stable Biotinylated Firefly Luciferase and Its Application in Pyrosequencing |
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169 | (10) |
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15 Expression of Thermostable Recombinant Luciola lateralis Luciferase and Its Function on Setting Up of Heat-Stable Pyrosequencing System |
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179 | (8) |
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16 Expression and Purification of ATP Sulfurylase from Saccharomyces cerevisias in Escherichia coli and Its Application in Pyrosequencing |
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187 | (10) |
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17 Characterization of Recombinant Escherichia coli Single-Strand Binding Protein and Its Application in Pyrosequencing |
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197 | (10) |
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18 Expression of PPDK from Microbispora rosea subsp. aerata in E. coli and Its Application in Pyrosequencing |
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207 | (10) |
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PART IV PYROSEQUENCING-BASED BIO-BARCODES |
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19 Construction of 3-Plex Barcodes for Differential Gene Expression Analysis with Pyrosequencing |
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217 | (14) |
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20 Development of Pyrosequencing-Based Multiplex Bioassay by Designing Barcodes Encoded with Artificially Designed Sequences |
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231 | (12) |
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21 Quantitatively Discriminating Multiplexed LAMP Products with Pyrosequencing-Based Bio-Barcodes |
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243 | (14) |
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22 Multiplex Ligation-Dependent Sequence-Tag Amplification Coupled with Pyrosequencing for Copy Number Variation (CNV) Analysis |
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257 | (10) |
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23 Pyrosequencer Miniaturized with Capillaries to Deliver Deoxynucleotides |
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267 | (10) |
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24 Pyrosequencing on Acryl-Modified Glass Chip |
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277 | (12) |
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25 Pyrosequencing On-Chip Based on a Gel-Based Solid-Phase Amplification |
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289 | (14) |
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PART VI CLINICAL PRACTICE |
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26 Prenatal Diagnosis of Trisomy 21 by Quantitatively Pyrosequencing Heterozygotes Using Amniotic Fluid as Starting Material of PCR |
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303 | (12) |
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27 Comparative Gene Expression Analysis of Breast Cancer-Related Genes by Multiplex Pyrosequencing Coupled with Sequence Barcodes |
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315 | (12) |
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28 MicroRNA Quantification by Pyrosequencing with a Sequence-Tagged Stem-Loop RT Primer |
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327 | (12) |
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29 Analysis of Genetically Modified Organisms by Pyrosequencing on a Portable Photodiode-Based Bioluminescence Sequencer |
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339 | (10) |
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30 Genotyping of Pathogenic Serotypes of S. suis with Pyrosequencing |
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349 | (12) |
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31 Differential Gene Expression Analysis of Breast Cancer by Combining Sequence-Tagged Reverse-Transcription PCR with Pyrosequencing |
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361 | (10) |
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32 Detection of Avian Influenza A Virus by Pyrosequencing |
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371 | (10) |
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33 Genotyping of Alcohol Dehydrogenase Gene by Pyrosequencing Coupled with Improved LATE-PCR Using Human Whole Blood as Starting Material |
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381 | (10) |
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34 Pyrosequencing on a Single Cell |
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391 | (8) |
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| Index |
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