DNA SequencingDNA sequencing is one of the most important techniques in molecular biology today. Everyone, at some time, has to sequence some DNA - whether just a few hundred base pairs, or a few megabases as part of a major sequencing project. Although it is widely practised, until now there did not exist a practical guide for the absolute beginner. Written in an accessible and informal style, this book answers the important questions: how does DNA sequencing work? What do you need to do before getting started? What does the jargon mean? What are all the reagents for? Why hasn't it worked? If you have ever asked any of these questions, DNA Sequencing: The Basics is for you. |
Contents
The sequencing reactions | 2 |
Further reading | 52 |
Reading the sequence from the autoradiograph | 65 |
Copyright | |
2 other sections not shown
Common terms and phrases
5'-triphosphate 7-deaza-dGTP acrylamide annealing autoradiograph bacteria bacteriophage bacterium banding pattern base pairing buffer carry cells chain terminated molecules chain termination method chain termination sequencing Chapter chemical degradation method chemical degradation sequencing cloned DNA cloning vector codons coli Databases ddATP ddNTPs deletions dideoxynucleotide dITP DNA fragments DNA polymerase DNA preparation DNA sequencing dNTPs double stranded DNA end-labelled enzyme exonuclease exonuclease activity Figure gel electrophoresis gel mix gene genome ghost bands hairpin loops infected inserted DNA IRL Press Klenow polymerase labelled nucleotide labelling step lacZ lane length M13 phage master sequence modified nucleotides molecular biology Nucleic acids nucleotides obtaining single stranded oligonucleotide overhang phage particles phagemid piperidine plaque plasmid plates polyacrylamide gel polymerized polynucleotide primer problem protein read the sequence region remove replication Running the gel Section Sequenase sequencing gel sequencing procedure sequencing reactions single stranded DNA solution strand synthesis reactions techniques template DNA tion triphosphate