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ABI Prism 310 Cycle Sequencing Protocol - http://vosshall.rockefeller.edu/protocols/ABICYCLESEQ.pdf

Category: DNA Protocols: 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]

Agarose Gel Electroporesis of DNA Protocol - http://www.unc.edu/~fbottone/protocol/electro.html

Category: Nucleic Acid Electrophoresis Protocols: DNA Electrophoresis Protocols: DNA Electrophoresis in Agarose Gels Protocols

Protocol for agarose gel electrophoresis of DNA. Includes: Making the gel; Loading the gel; Running the gel.
- [Read Agarose Gel Electroporesis of DNA Protocol]

Calcium Flux: Indo-1 Loading and Sample Staining Procedure for Simultaneous Measurement of Intra Ca2 - http://www.natureprotocols.com/2007/01/29/calcium_flux_indo1_loading_and.php

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Intracellular Ion Measurement Protocols

Protocol for Calcium Flux: Indo-1 loading and sample staining procedure for simultaneous measurement of intracellular Ca2+. Includes: Preparation of INDO-1; Preparation of Ionomycin. - [Read Calcium Flux: Indo-1 Loading and Sample Staining Procedure for Simultaneous Measurement of Intra Ca2]

Cell Fractionation of a Mixed Population of Cells - http://www.axis-shield.com/densityhome/optiprep/C14.pdf

Category: Cell Biology and Cell Culture

Cell fractionation techniques are presented for the design of gradient systems for separating one or more cell types from lavages of body cavities or from mechanically or enzymically-dissociated tissues. Includes: Preparation of cell suspension for gradient loading; Fractionation by buoyant density; Fractionation on the basis of cell size. - [Read Cell Fractionation of a Mixed Population of Cells]

DNA Preparation and Needle Loading for Gene Delivery by Microinjection Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4652

Category: Microinjection Protocols: Microinjection of DNA Protocols

For microinjection into cells, high-quality plasmid DNA is purified using standard procedures. The resulting preparation is then delivered into the microinjection needle by either a back-filling or a forward-filling approach. - [Read DNA Preparation and Needle Loading for Gene Delivery by Microinjection Protocol]

DNA Sequencing Methods - http://www.genome.ou.edu/protocol_book/protocol_partIV.html#IV.C

Category: DNA Protocols: DNA Sequencing Protocols

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]

Dot and Slot Hybridization of Purified RNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4052

Category: Nucleic Acid Detection Protocols: Nucleic Acid Blotting Protocols

Protocol for dot and slot hybridization of purified RNA. Dot blotting of RNA is best carried out using purified preparations of RNA that are denatured with glyoxal or formaldehyde immediately before loading onto a nylon membrane through a vacuum manifold. - [Read Dot and Slot Hybridization of Purified RNA Protocol]

General Method for Ras, Rac, Cdc42, and Rho Activation Assay - http://www.upstate.com/misc/protocol_detail.q.prot.e.general_method_ras.a.name.e.General_Method_for_Ras_Rac_Cdc42_and_Rho_Activation_Assay

Category: Signalling Signal Transduction Protocols

General protocol for Ras, Rac, Cdc42, and Rho activation assay. Includes: Affinity Precipitation/Immunoblot Protocol, Cell Culture and Extract Preparation (Adherent and Non Adherent cells), GTPÎ³S/GDP Loading for Positive and Negative Controls, Ras, Rac ,Cdc42, and Rho Pull-Down Assay and Western Blot and Detection. - [Read General Method for Ras, Rac, Cdc42, and Rho Activation Assay]

Indo-1 loading and sample staining procedure for simultaneous measurement of intracellular Ca2+ - http://www.natureprotocols.com/2007/01/29/calcium_flux_indo1_loading_and.php

Category: Signalling Signal Transduction Protocols: Calcium Measurement Protocols

Protocol by using the technique of Indo-1 loading and sample staining procedure for simultaneous measurement of intracellular Ca2+. Protocol includes: Preparation of Indo-1 and preparation of Ionomycin. - [Read Indo-1 loading and sample staining procedure for simultaneous measurement of intracellular Ca2+]

Loading and Running DNA Sequencing Gels Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3794

Category: DNA Protocols: DNA Sequencing Protocols

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

Measurement of Intracellular Ions by Flow Cytometry Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E661C4C9AC9935B31A1C2087E2276A3&objectid=6674EF32EE59A37C48B4A267D733CAE2

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Intracellular Ion Measurement Protocols

Describes flow cytometric protocols using the dyes Indo-1 AM, Fluo-3, and Fura Red AM to measure intracellular calcium concentration. Support protocols detail the use of calcium buffers to calibrate a flow cytometric calcium assay, and methods to facilitate dye loading; an alternate protocol describes the use of a spectrofluorimeter to measure intracellular calcium for those investigators without access to a flow cytometer. - [Read Measurement of Intracellular Ions by Flow Cytometry Protocol]

Membrane Potential Analysis by Flow Cytometry Protoocol - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/membrane.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for membrane potential analysis by flow cytometry. Includes: Preparation of the dyes; Loading of the dyes; Flow Cytometric Analysis. - [Read Membrane Potential Analysis by Flow Cytometry Protoocol]

Microinjecting C.elegans Worms Protocol - http://cobweb.dartmouth.edu/cgi-bin/cgiwrap/~ambros/protocol.cgi?id=5

Category: C. elegans Protocols: C. elegans Injection Protocols

Protocol for microingecting C.elegans worms. Includes: Making the DNA solution; Setting up the scope, loading the needle, mounting, and breaking the needle; Mounting worms on an injection pad; Injecting. - [Read Microinjecting C.elegans Worms Protocol]

Preparation and Loading of Protein Samples for Microinjection Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4656

Category: Microinjection Protocols: Microinjection of Proteins Protocols

his protocol provides methods for the preparation of protein samples and for loading them into pulled microinjection pipettes. Stock solutions of proteins are thawed, diluted (if desired), centrifuged at high speed to remove aggregates, and kept on ice until loading. Loading into micropipettes can be done using either a "front-loading" or a "backfilling" procedure. - [Read Preparation and Loading of Protein Samples for Microinjection Protocol]

Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of DNA from Mammalian Cells - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4030

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol for preparation of DNA for pulsed-field gel electrophoresis: isolation of DNA from mammalian cells and tissues. Genomic DNAs from mammalian cells are prepared for pulsed-field gel electrophoresis by lysing cells in situ in an agarose plug. Following digestion with an appropriate restriction enzyme, the plug is loaded directly into the well of a pulsed-field gel or it can be melted before loading. - [Read Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of DNA from Mammalian Cells]

Protocol for Microtubule Binding Assays - http://www.proweb.org/kinesin/Methods/MT-binding_assay.html

Category: Cell Biology and Cell Culture

Microtubule binding assay protocol using the materials: Siliconized ultracentrifuge microfuge tubes, GTP-depleted microtubules, 6X SDS loading dye, 1X SDS loading dye, Coomassie Brilliant Blue R 250 (0.8% in 50% methanol + 10% acetic acid)and Destaining solution (13% methanol + 3% acetic acid). - [Read Protocol for Microtubule Binding Assays]

PROTOCOL FOR SDS-PAGE - http://www.fiu.edu/~animals/protocols/sds_page.html

Category: Protein Protocols: Protein Electrophoresis: SDS-PAGE Electrophoresis Protocols

PROTOCOL FOR SDS-PAGE. Immuno-Physiology Laboratory, Florida International University. Includes: Primary considerations, preparing samples, loading samples, running gels, and quantitative staining. - [Read PROTOCOL FOR SDS-PAGE]

Purification of viable bovine spermatozoa of normal morphology. - http://www.axis-shield.com/densityhome/optiprep/C16.pdf

Category: Cell Biology and Cell Culture

Protocol for the recovery of a highly viable fraction for use in fertilization. Protocol uses an iodinated density gradient medium to adjust the density of the raw ejaculate to approx 1.170 g/ml by mixing with a high density medium (Ï >1.26 g/ml) and loading it beneath a discontinuous gradient. - [Read Purification of viable bovine spermatozoa of normal morphology.]

Simultaneous Measurement of Intracellular CA2+ and Cell Surface Antigen Expression Protocol - http://cyto.mednet.ucla.edu/Protocols/Calcium%20Staining.htm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Intracellular Ion Measurement Protocols

INDO-1 loading and sample staining protocol for simultaneous measurement of intracellular CA2+ and cell surface antigen expression. Includes: Preparation of INDO-1; Preparation of Ionomycin. - [Read Simultaneous Measurement of Intracellular CA2+ and Cell Surface Antigen Expression Protocol]

Sodium and Potassium Analysis by Flow Cytometry Protocol - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/sodium.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

Protocol for sodium and potassium analysis by flow cytometry. Includes: Preparation of the dyes; Loading of the dyes; Flow Cytometric Analysis. - [Read Sodium and Potassium Analysis by Flow Cytometry Protocol]

Sodium and Potassium Analysis by Flow Cytometry Protocol - http://www.niehs.nih.gov/research/atniehs/core/fc/protocols/sodium.cfm

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Intracellular Ion Measurement Protocols

Articles (1-4 of 4)

Quick DNA Agarose Gel Electrophoresis Protocol.

A quick and simple protocol for the electrophoresis of DNA fragments using agarose gels.

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.