Biotechnology procedures and experiments handbook

[...]... net charge, size and shape of the molecules, and also on the ionic strength, viscosity, and temperature of the medium in which the molecules are moving As an analytical tool, electrophoresis is simple, rapid, and highly sensitive It is used analytically to study the properties of a single charged species, and as a separation technique 14 BIOTECHNOLOGY PROCEDURES AND EXPERIMENTS HANDBOOK There are... ammonium acetate, 0.3 M for sodium acetate, 0.2 M for sodium chloride, and 0.8 M for lithium chloride The ion used often depends on the volume of DNA and the subsequent manipulations; for 8 BIOTECHNOLOGY PROCEDURES AND EXPERIMENTS HANDBOOK example, sodium acetate inhibits Klenow, ammonium ions inhibit T4 polynucleotide kinase, and chloride ions inhibit RNA-dependent DNA polymerases The addition of... Composition of all buffers should be 1 2 BIOTECHNOLOGY PROCEDURES AND EXPERIMENTS HANDBOOK included, unless they are standard or included in a kit Include all calculations made in preparing solutions Biological reagents should be identified by their original source, for example, genotype, for strains; concentration, source, purity, and/ or restriction map for nucleic acids; and base sequence, for oligonucleotides... molecular weight standards Measure the distances of each unknown band For samples lanes with many bands (serum in this exercise), measure all bands in those with just a few and the major bands in those that have many Prepare a standard curve by plotting log MW versus relative mobility (Rf, distance traveled by protein divided by distance traveled by dye) Use this and the mobility of bands from your fractions... concentration of the light-absorbing substance Method 1 Turn on the spectrophotometer and allow 10 minutes for the instrument to warm up before use 2 Adjust the wavelength to that specified for the procedure you are using 12 BIOTECHNOLOGY PROCEDURES AND EXPERIMENTS HANDBOOK 3 Be sure the cover is closed on the cuvette holder and use the left knob on the front panel to adjust the dark current so that the... mark the backs of the plates with your initials, the date, and relevant experimental data, e.g., strain numbers Preparation of Solutions (a) Calculation of Molar, %, and "X" Solutions (i) A molar solution is one in which 1 liter of solution contains the number of grams equal to its molecular weight 4 BIOTECHNOLOGY PROCEDURES AND EXPERIMENTS HANDBOOK Example To make up 100 mL of a 5M NaCl solution =... spectrophotometer differs from a calorimeter in the manner in which light is 9 10 BIOTECHNOLOGY PROCEDURES AND EXPERIMENTS HANDBOOK separated into its component wavelengths A spectrophotometer uses a prism to separate light and a calorimeter uses filters Both are based on a simple design, passing light of a known wavelength through a sample and measuring the amount of light energy that is transmitted This is accomplished... reagents and conditions are important considerations in processing and storing DNA and RNA Heavy metals promote phosphodiester breakage EDTA is an excellent heavy metal chelator Free radicals are formed from chemical breakdown and radiation and they cause phosphodiester breakage UV light at 260 nm causes a variety GENERAL I NSTRUCTION AND LABORATORY METHODS 7 of lesions, including thymine dimers and crosslinks... polymerized, the acrylamide between the plates will also be ready 16 BIOTECHNOLOGY PROCEDURES AND EXPERIMENTS HANDBOOK 4 If you are running the gel on the same day, prepare samples while the acrylamide is polymerizing Otherwise, wait until you are ready to run the gel (i) You will need a sample of each unknown substance, plus the molecular weight standards Prepare samples in screw-cap microcentrifuge tubes The... Procedure: Write down the exact procedure and flow chart before you perform each experiment, and make sure you understand each step before you do it You should include everything you do, including all volumes and amounts; many protocols are written for general use and must be adapted for a specific application Writing a procedure helps you to remember and understand what it is about It will also help . [Introduction to practical biotechnology] Biotechnology procedures and experiments handbook / S. Harisha. p. cm. — (An introduction to biotechnology) Originally. be Chapter 1 GENERAL INSTRUCTION AND LABORATORY METHODS 1 2 BIOTECHNOLOGY PROCEDURES AND EXPERIMENTS HANDBOOK included, unless they are standard or included in a