knowledge Protocols

Category: Microscopy Protocols: Live-Cell Imaging Protocols

When choosing a particular molecule for photoactivation studies, it is necessary to have some structural knowledge of the molecule in order to design an appropriately caged species that will retain its biological inactivity until uncaging is effected. Includes synthesis of caged peptides or proteins. - [Read Design, Synthesis, and Characterization of Caged Compounds]

Category: Microinjection Protocols: Microinjection of Proteins Protocols

When choosing a particular molecule for photoactivation studies, it is necessary to have some structural knowledge of the molecule in order to design an appropriately caged species that will retain its biological inactivity until uncaging is effected. - [Read Design, Synthesis, and Characterization of Caged Compounds Protocol]

Category: DNA Protocols: DNA Extraction Protocols: DNA Extraction Polyacrylamide Gels

DNA Electroelution.  This protocol describes the purification of DNA by trapping in a high-salt cushion in a "UEA AnalyticalElectroeluter" (IBI).  This machine is no longer manufactured, to our knowledge.  However, a smiliar device can be easily made from Plexiglas according to the following diagram, taken from Cornel Mulhardt, Molecular Biology and Genomics (2007) Academic Press, p.52: Schimenti Lab - [Read DNA Electroelution]

Category: PCR Protocols: cDNA Synthesis Protocols

3'-RACE reactions are used to isolate unknown 3' sequences or to map the 3' termini of mRNAs onto a gene sequence.  3'-RACE requires knowledge of a small region of sequence within either the target RNA or a partial clone of cDNA.  A population of mRNAs is transcribed into cDNA with an adaptor-primer consisting at its 3' end of a poly(T) tract and at its 5' end of an arbitrary sequence of 30-40 nucleotides. - [Read Rapid Amplification of 3' cDNA Ends 3'-RACE Protocol]

Category: PCR Protocols: cDNA Synthesis Protocols

This method is used to extend partial cDNA clones by amplifying the 5' sequences of the corresponding mRNAs.  The technique requires knowledge of a small region of sequence within the partial cDNA clone.  During PCR, the thermostable DNA polymerase is directed to the appropriate target RNA by a single primer derived from the region of known sequence. - [Read Rapid Amplification of 5' cDNA Ends 5'-RACE Protocol]

Category: Chromatography Protocols: Ion Exchange Chromatography Protocols

Protocol for selection of an ion exchanger: Determining the pI of a protein using isoelectric focusing. The choice of whether to use an anion or a cation exchanger should be based on knowledge of the stability of the protein, and the binding properties of the target protein and other molecules present. - [Read Selection of an Ion Exchanger Protocol]

Category: Chromatography Protocols: Ion Exchange Chromatography Protocols

Protocol for selection of an ion exchanger: Determining the pI of a protein using the titration curve method. The choice of whether to use an anion or a cation exchanger should be based on knowledge of the stability of the protein, and the binding properties of the target protein and other molecules present in the sample. - [Read Selection of an Ion Exchanger Protocol II]

Category: Chromatography Protocols: Ion Exchange Chromatography Protocols

Protocol for selection of an ion exchanger: Determining the pI of a protein using the test tube method. The choice of whether to use an anion or a cation exchanger should be based on knowledge of the stability of the protein, and the binding properties of the target protein and other molecules present in the sample. - [Read Selection of an Ion Exchanger Protocol III]

Category: Chromatography Protocols: Ion Exchange Chromatography Protocols

Protocol for selection of an ion exchanger: Determining the pI of a protein using the Trial-and-Error method. The choice of whether to use an anion or a cation exchanger should be based on knowledge of the stability of the protein, and the binding properties of the target protein and other molecules present in the sample. - [Read Selection of an Ion Exchanger Protocol IV]