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Large Scale Tubulin Preparation

Tubulin is purified from bovine/porcine brain by two cycles of polymerization/depolymerization followed by removal of copurifying proteins on a phosphocellulose (PC) column. The procedure described he ...

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Ethidium Bromide Decontamination

N.B.: Ethidium bromide is a powerful mutagen. Protective gloves should be worn at all times when handling solutions containing ethidium bromide. Decontamination of solutions 0.5mg/mlThis method reduc ...

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Appendix F: Centrifugation

The single most important advance in the use of centrifugal force to separate biologically important substances. was the coupling of mechanics optics and mathematics by T. Svedberg and J.W. Williams i ...

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Thin Layer Chromatography

Reaction volume 100 ul5 ul NADPH (100 mM)x ul cell extract so you get 50 % conversion0.2 uM C14-Testosteroneadd Tris-Citrate Buffer pH 6 to an endvolume of 100 ulmix C14-Testosterone and Citrate Buffe ...

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Detection and Measurement of Radioactivity

Radioactive DecayIsotopes of a given element have nuclei with the same number of protons but different numbers of neutrons. Some isotopes are stable however radioisotopes are unstable and disintegrate ...

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Quantification made easy

With the aid of spectroscopy the quantitative analysis of nucleic acids and proteins has established itself as a routine method in many laboratories. It includes absorption measurements in the ultravi ...

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Appendix G: Spectrophotometry

Figure G.1 Electromagnetic Radiation Spectrum Figure G.2. Schematic light transmission Figure G.3. Use of the Spec 20 A spectrophotometer or colorimeter makes use of the transmission of light through ...

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A. Gilson-style Pipetmen Micropipets (and similar models)

We primarily use the Gilson micropipets in the core course labs. We have four sizes identified by the number on the round button on the plunger. The value is the maximum volume in microliters that can ...

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How to prepare Molecular Biology grade glycogen

OverviewGlycogen can conveniently substitute for tRNA as a carrier for nucleic acid precipitation. Although Molecular Biology grade glycogen can be purchased from a number of vendors the main disadva ...

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Marcantonio Lab Protocol Manual

Alkali DenaturationAlkali Denaturation of Supercoiled Plasmid DNA 1) To 20 ul of purified concentrated DNA solution add 2 ul of freshly prepared NaOH/EDTA solution and incubate at 37 degrees for 30 mi ...

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Preparation of Sonicated Human DNA

Purpose: To break up high molecular weight human placental DNA into fragment sizes of 500 bp or less which can be used as competitor DNA in Southern and in situ hybridizations. This method can also be ...

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A protocol for cleaning and reusing the large 25 x 25 cm plates

We regularly reuse our large 25x25 cm plating trays; initially however we were plagued by gross microbiological contamination when reusing the trays. Various combinations of cleaning with a laboratory ...

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Microbiology - Serial Dilution

1. An undiluted (stock) solution is termed the 100 dilution. This may be a chemical solution or a suspension of bacteria or bacteriophage.2. Typical diluions are 10-fold dilutions; each dilution is 1/ ...

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Silane and Poly-L-Lysine Coating of Microscope Slides

1) Wash slides in Decon solution for 30 minutes. 2) Wash slides in running tap water for 30 minutes. 3) Wash slides in distilled water for 2 x 5 minutes. 4) Wash slides in 95% ethanol for 2 x 5 minute ...

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Siliconizing Glassware

Caution: Wear gloves and lab coat. Work with dichlorodimethylsilene in hood. Fill large dessicator with glassware. Put 1 ml dichlorodimethylsilene in a small beaker and put it in the dessicator. Pull ...

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Aseptic Techniques

Aseptic techniques ensure that all cell culture procedures are performed to a standard that will prevent contamination from bacteria fungi mycoplasma and cross contamination with other cell lines. All ...

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Asepsis and Aseptic Practices in the Operating Room

Preventing surgical site infection in the operating room is the primary goal of the surgical team and all activities performed by the team support this goal. Some of these activities include patient r ...

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Formaldehyde Treatment of Tissue Culture Hoods

You will need:- 12g Potassium Permanganate 6g Crushed paraformaldehyde 1) Set the hood so that it can vent outside. 2) Mix the two chemicals above together (in a jam/coffee jar so it can be thrown awa ...

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Sterile Technique

Good sterile technique is the first and most important step in insuring consistent results when employing recombinant DNA and protein expression techniques. Sterile technique refers to procedures by w ...

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DEPC

Procurement100ml DEPC from Sigma (D5758) - �33/$292 (as of 2009-01) 100ml DEPC from MPI (150902) - �13 (as of 2009-01) DEPC-treatment of solutionsadd 0.05-0.1% (v/v) DEPC to solution/water (More DEP ...

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