Advances and Clinical Practice in Pyrosequencing

Part I: Template Preparation

1. Preparation of Single-Stranded DNA for Pyrosequencing by LATE-PCR

Huiyong Yang, Chao Liang, Zhiyao Chen, Bingjie Zou, Qinxin Song, and Guohua Zhou

2. A Simplified Protocol for Preparing Pyrosequencing Templates Based on LATE-PCR Using Whole Blood as Starting Material Directly

Yunlong Liu, Haiping Wu, Hui Ye, Zhiyao Chen, Qinxin Song, Bingjie Zou, Jianzhong Rui, and Guohua Zhou

3. Improvement of LATE-PCR to Prepare Pyrosequencing Template

Qinxin Song, Huiyong Yang, Bingjie Zou, Hideki Kambara, and Guohua Zhou

4. Pyrosequencing Templates Generated by Nicking PCR Products with Nicking Endonuclease

Qinxin Song, Haiping Wu, Guohua Zhou, and Hideki Kambara

5. Pyrosequencing Templates Generated by Asymmetric Nucleic Acid Sequence-Based Amplification (Asymmetric-NASBA)

Huning Jia, Zhiyao Chen, Haiping Wu, Hui Ye, Bingjie Zou, Qinxin Song, and Guohua Zhou

6. Gel Immobilization of Acrylamide-Modified Single-Stranded DNA Template for Pyrosequencing

Pengfeng Xiao, Huan Huang, Bingjie Zou, Qinxin Song, Guohua Zhou, and Zuhong Lu

7. Multiplex PCR Based on a Universal Biotinylated Primer to Generate Templates for Pyrosequencing

Zhiyao Chen, Yunlong Liu, Hui Ye, Haiping Wu, Jinheng Li, Bingjie Zou, Qinxin Song, and Guohua Zhou

Part II: Technology Innovation

8. A Novel Pyrosequencing Principle Based on AMP-PPDK Reaction for Improving the Detection Limit

9. Pyrosequencing Chemistry Coupled with Modified Primer Extension Reactions for Quantitative Detection of Allele Frequency in a Pooled Sample

Guohua Zhou, Masao Kamahori, Kazunori Okano, Gao Chuan, Kunio Harada, and Hideki Kambara

10. A Gel-Free SNP Genotyping Method Based on Pyrosequencing Chemistry Coupled with Modified Primer Extension Reactions Directly from Double-Stranded PCR Products

Guohua Zhou, Hiromi Shirakura, Masao Kamahori, Kazunori Okano, Keiichi Nagai, and Hideki Kambara

11. Improvement of Pyrosequencing to Allow Multiplex SNP Typing in a Pyrogram

Guohua Zhou, Mari Gotou, Tomoharu Kajiyama, and Hideki Kambara

12. Improvement of P

Haiping Wu, Wenjuan Wu, Zhiyao Chen, Weipeng Wang, Bingjie Zou, Qinxin Song, Guohua Zhou, and Hideki Kambara

13. Using Polymerase Preference Index to Design imLATE-PCR Primers for an Efficient Pyrosequencing

Qinxin Song, Xiemin Qi, Yiyun Shen, Yiru Jin, Bingjie Zou, and Guohua Zhou

Part III: Reagents Preparation

14. Preparation of Thermalstable Biotinlyted Firefly Luciferase and Its Application in Pyrosequencing

Shuhui Zhu, Bingjie Zou, Haiping Wu, Yinjiao Ma, Ying Chen, Qinxin Song, and Guohua Zhou

15. Expression of Thermostable Recombinant Luciola lateralis Luciferase and Its Function on Setting up of Heat-Stable Pyrosequencing System

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16. Expression and Purification of ATP Sulfurylase from Saccharomyces cerevisias in Escherichia coli and Its App