𝔖 Bobbio Scriptorium
✦   LIBER   ✦

Improved Performance of Pyrosequencing Using Single-Stranded DNA-Binding Protein

✍ Scribed by Mostafa Ronaghi

Publisher
Elsevier Science
Year
2000
Tongue
English
Weight
102 KB
Volume
286
Category
Article
ISSN
0003-2697

No coin nor oath required. For personal study only.

✦ Synopsis

In modern biology, there is a critical need to develop a high-throughput and inexpensive platform for DNA sequencing. Pyrosequencing is a nonelectrophoretic single-tube DNA sequencing method that takes advantage of cooperativity between four enzymes to monitor DNA synthesis. In these studies, single-stranded DNAbinding protein (SSB) was added to the primed DNA template prior to the Pyrosequencing reaction. The addition of SSB to a Pyrosequencing reaction system resulted in a read length of more than 30 nucleotides. Improvements were observed as: (i) increased efficiency of the enzymes, (ii) reduced mispriming, as measured by nonspecific signals, (iii) an increase in signal intensity during the reaction, (iv) higher accuracy in reading the number of identical adjacent nucleotides in difficult templates, and (v) longer reads. The usefulness of these results for future Pyrosequencing applications is discussed.

📜 SIMILAR VOLUMES

Escherichia coli single-stranded DNA-bin
Escherichia coli single-stranded DNA-binding protein, a molecular tool for improved sequence quality in pyrosequencing
✍ Maria Ehn; Afshin Ahmadian; Peter Nilsson; Joakim Lundeberg; Sophia Hober 📂 Article 📅 2002 🏛 John Wiley and Sons 🌐 English ⚖ 207 KB 👁 1 views

Pyrosequencing is a four-enzyme bioluminometric DNA sequencing technique based on a DNA sequencing by synthesis principle. Currently, the technique is limited to analysis of short DNA sequences exemplified by single-nucleotide polymorphism analysis. In order to expand the field for pyrosequencing, t

Fast detection of quadruplex structure i
Fast detection of quadruplex structure in DNA by the intrinsic fluorescence of a single-stranded DNA binding protein
✍ Xin-ying Zhuang; Jun Tang; Yu-hua Hao; Zheng Tan 📂 Article 📅 2007 🏛 John Wiley and Sons 🌐 English ⚖ 300 KB 👁 1 views

## Abstract Single‐stranded guanine‐rich (G‐rich) DNA can fold into a four‐stranded G‐quadruplex structure and such structures are implicated in important biological processes and therapeutic applications. So far, bioinformatic analysis has identified up to several hundred thousand of putative quad

Tetraplex formation of surface-immobiliz
Tetraplex formation of surface-immobilized human telomere sequence probed by surface plasmon resonance using single-stranded DNA binding protein
✍ Zhi-Xiong Zeng; Yong Zhao; Yu-Hua Hao; Zheng Tan 📂 Article 📅 2005 🏛 John Wiley and Sons 🌐 English ⚖ 134 KB

Many sequences in genomic DNA are able to form unique tetraplex structures. Such structures are involved in a variety of important cellular processes and are emerging as a new class of therapeutic targets for cancers and other diseases. Screening for molecules targeting the tetraplex structure has b

The effects of combined antioxidant (β-c
The effects of combined antioxidant (β-carotene, α-tocopherol and ascorbic acid) supplementation on antioxidant capacity, DNA single-strand breaks and levels of insulin-like growth factor-1/IGF-binding protein 3 in the ferret model of lung cancer
✍ Yuri Kim; Fuzhi Lian; Kyung-Jin Yeum; Nalinee Chongviriyaphan; Sang-Woon Choi; R 📂 Article 📅 2007 🏛 John Wiley and Sons 🌐 French ⚖ 254 KB 👁 1 views

## Abstract Insulin‐like growth factor 1 (IGF‐1) and its major binding protein, IGF binding protein 3 (IGFBP‐3) are implicated in lung cancer and other malignancies. We have previously shown that the combination of three major antioxidants [β‐carotene (BC), α‐tocopherol (AT) and ascorbic acid (AA)]