DNA Sequencing Protocols

Protocol for ABI prism 310 cycle sequencing. Includes: Sequencing Reaction; Column Purification; Sample Loading Preparation. - [Read ABI Prism 310 Cycle Sequencing Protocol]

Protocol describes fixation of a sequencing gel and processing in preparation for autoradiography. - [Read Autoradiography and Reading of Sequencing Gels Protocol]

Protocol for chemical sequencing. - [Read Chemical Sequencing Protocol]

Protocol first describes the vector preparation and, then, describes the insert preparation.  Vital to have an excellent vector in order to produce a sequencing library.  Protocol employs the male-specific coliphage M13 as the sequencing vector.  M13 is a filamentous phage with a single-stranded, circular genome.  M13 is widely used as a vector because many versions are available commercially and because M13 has certain advantages. - [Read Construction of the Sequencing Library Protocol]

Protocol for cycle sequence reactions for large insert plasmid templates. Includes: CycleSequenceConditions; Primers Useful For BAC and PAC Vectors. - [Read Cycle Sequence Reactions For Large Insert Plasmid Templates Protocol]

Protocol for cycle sequencing for PCR product. - [Read Cycle Sequencing for PCR Product Protocol]

Direct sequencing from amplified bacterial and large insert cloned human or mouse genomic DNA via an improved MultiPlex PCR-based method. Includes: Preparing primers for MP-PCR; Amplification; PCR Product Clean-up; Sequencing. - [Read Direct Sequencing Using MultiPlex PCR-Based Method]

Methods for DNA sequencing.  Includes: Bst-catalyzed radiolabeled DNA sequencing; Radiolabeled sequencing gel preparation, loading, and electrophoresis; Taq-polymerase catalyzed cycle sequencing using fluorescent-labeled dye primers; Taq-polymerase catalyzed cycle sequencing using fluorescent-labeled dye terminator reactions; Sequenase[TM] catalyzed sequencing with dye-labeled terminators; etc. - [Read DNA Sequencing Methods]

Protocol describes a simple procedure for purifying sequencing reactions using the MagneSil particles from Promega. - [Read Dye Terminator Cleanup for Automated DNA Sequencing Reactions Protocol]

Protocol for loading and running DNA sequencing gels. - [Read Loading and Running DNA Sequencing Gels Protocol]

Protocol for Maxam- Gilbert sequencing. - [Read Maxam-Gilbert Sequencing Protocol]

Protocol for plasmid sequencing. - [Read Plasmid Sequencing Protocol]

The inclusion of formamide in sequencing gels eliminates secondary structure in the DNA during electrophoresis. Formamide gels are particularly useful and almost a necessity when sequencing DNA templates with a G/C content >55%. - [Read Preparation of Denaturing Polyacrylamide Gels Containing Formamide Protocol]

Protocol for sequencing difficult regions on DNA. - [Read Sequencing Difficult DNA Regions Protocol]

Protocol for single strand DNA prep for sequencing. Protocol works well from a 5 ml starting culture or a 1 ml starting culture. - [Read Single Strand DNA Prep for Sequencing Protocol]