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The Q5 High-Fidelity 2X Master Mix offers robust, high-fidelity performance in a convenient master mix format. The Q5 High- Fidelity 2X Master Mix features a high-fidelity, thermostable DNA polymerase with 3´→ 5´ exonuclease activity, fused to a processivity-enhancing Sso7d domain to support robust DNA amplification. With an error rate ~280-fold lower than that of Taq DNA Polymerase, Q5 High-Fidelity DNA Polymerase is ideal for cloning and can be used for long or difficult amplicons. The convenient master mix formulation is supplied at a 2X concentration. The mix contains dNTPs, Mg++ and a proprietary broad-use buffer requiring only the addition of primers and DNA template for robust amplification regardless of GC content. When used at the recommended 1X final concentration, the Q5 High-Fidelity Master Mix contains 2 mM Mg++. Q5 High-Fidelity DNA Polymerase is unlike typical, lower fidelity PCR enzymes. To determine the optimal annealing temperatures for a given set of primers, use of the NEB TmCalculator is highly recommended.

2X PCR Master Mixes are convenient to use as they include all the necessary PCR components except the template and primers. Most PCR Master Mixes also include agarose gel running dyes and a density reagent that allows direct loading of PCR products on an agarose gel for electrophoresis. The master mix format simplifies workflows and sample handling; simply add primers, template, and water and then begin PCR.

SapphireAmp Fast PCR Master Mix contains a hot start PCR enzyme, optimized buffer, dNTP mixture, gel loading dye (blue), and a density reagent as a 2X premix. SapphireAmp Fast PCR Master Mix is optimized for fast PCR and offers a rapid extension rate (10 sec. per kb). The inclusion of blue dye and a density reagent allows direct loading of PCR products on an agarose gel for electrophoresis. The master mix format simplifies workflows and sample handling; simply add primers, template, and water and then begin PCR. SapphireAmp Fast PCR Master Mix is ideal for fast colony PCR screening. Fast colony PCR amplification of a 5 kb insert can be completed in approximately 1 hr 15 min. Furthermore, it is possible to amplify fragments up to 6 kb from genomic DNA templates.

Prepare and run your SDS-PAGE gel using the Bio-Rad TGX gel system and the mini-PROTEAN gel box.

Working alone in the laboratory should be avoided; however, trained personnel can work alone in the laboratory if they adhere to the following Standard Operating Procedure (SOP). Students in training (during at least the 1st semester of laboratory work) cannot work alone in the laboratory.

The Thermo Scientific HisPur Ni-NTA Resin enables effective immobilized metal affinity chromatography (IMAC) purification of polyhistidine-tagged proteins from a soluble protein extract. This resin is composed of nickel-charged nitrilotriacetic acid (NTA) chelate immobilized onto 6% crosslinked agarose. The Ni-NTA resin is compatible with native or denaturing conditions and can be used in multiple formats, including conventional gravity-flow chromatography, spin column and FPLC. Ni-NTA resins are commonly chosen for His-tagged-protein purification because of the four metal-binding sites on the chelate, which allow for high-binding capacity and low-metal ion leaching.

This protocol is modified from the Drop Dialysis protocol available at http://www.neb.com. Introduction Many enzymes are adversely affected by a variety of residues in typical DNA preparations (e.g. minipreps, genomic and CsCl2 preparations). The following method has been successfully used to remove inhibitory substances (e.g. SDS or excess salt) from substrates intended for subsequent DNA manipulations. It is particularly effective for: Preparation of templates for DNA sequencing Assuring complete cleavage of DNA by restriction endonucleases Increasing the efficiency of ligation Reference 1. Silhavy, T., Berman, M. and Enquist, L. Experiments with Gene Fusions, Cold Spring Harbor, N.Y. Press (1984).

Will you be needed as a teacher in the year 2000? Should your school board buy land now for future expansion, or should it sell land already held? Would it be a good idea for an investor to put money into child-related industries for a maximum re turn twenty years from now? Answers to such questions depend at least partly on how many children will be around in the year 2000. Will your retirement fund be solvent when you retire? Will our nation's social security system be broke by the time you are eligible for benefits? What kinds of pro grams should churches and social agencies gear toward for maximum impact twenty years from now? Will we have enough space for people to live comfortably in the year 2000? What will we eat? How will we heat our homes and run our factories? To begin to answer any of these questions, fairly good estimates of the population lev els for the remainder of the century are needed by educators, urban planners, ecol ogists, scientists, and others. Population growth trends can be ana lyzed mathematically and future popu lation levels projected. Good projections on future population levels in the United States can be obtained using secondary school mathematics; thus population mod eling can be handled by secondary school students. In this article we will discuss the major factors affecting population change. A model will be developed that will allow us to start with the 1970 population figures, broken down by sex and age, and make projections at five-year intervals from 1975 to 2000. The principal sources of data are the Statistical Abstract of the United States and the Vital Statistics of the United States, both of which are published by the federal government and are available in most li braries. Much of the data can also be found in popular almanacs. Various factors influence population growth. The number of people immigrating into the United States increases the popu lation, as does the number of babies born. The number of people leaving the country and the number of individuals dying de crease the population. The roles of these factors may vary from year to year. The food supply and birth control programs may change the birth and survival rates over a span of time. Government regula tions may alter immigration laws. New dis coveries in medicine and nutrition may al ter life expectancy, and natural disasters and epidemics will change the death rate. Catastrophes, such as earthquakes, epi demics, and wars will influence population levels, but since there is no good way to predict catastrophes, most models ignore them in making projections. The major components of population change are birth, death, immigration, and emigration. We shall give a brief ex