Recent technological advances have enabled visualization of the organization and dynamics of local �chromatin structures; however, the global mechanisms by which chromatin organization modulates gene regulation are poorly understood. We designed and constructed a human a ...

DNA-based transposons are natural gene delivery vehicles. Similarly to retroviruses, these elements �integrate into the chromosomes of host cells, but their life-cycle does not involve reverse transcription and they are not infectious. Transposon-based gene delivery has seve ...

Mammalian artificial chromosomes (MACs) are engineered chromosomes with defined genetic content that can function as non-integrating vectors with large carrying capacity and stability. The large carrying capacity allows the engineering of MACs with multiple copies of the same ...

Advances in mammalian artificial chromosome technology have made chromosome-based vector technology amenable to a variety of biotechnology applications including cellular protein production, genomics, and animal transgenesis. A pivotal aspect of this technology is the a ...

The science of the so-called polymerase chain reaction (PCR) is now well known. However, the legal story associated with PCR is, for the most part, not understood and constantly changing. This presents diffculties for scientists, whether in academia or industry, who wish to practice the PCR proc ...

The development of the polymerase chain reaction (PCR) has often been likened to the development of the Internet, and although this does risk overstating the impact of PCR outside the scientific community, the comparison works well on a number of levels. Both inventions have emerged in the last 20 y ...

Polymerase chain reaction (PCR) is a very sensitive method of amplifying specific nucleic acid, but the system is susceptible to contamination from extraneous or previously amplified DNA strands (1,2). Many specific copies of DNA are produced from each round of amplification (3) with a sing ...

RNA extraction is fundamental to all aspects of mRNA analysis. We include here a simple method that avoids the use of a mortar and pestle.

Polymerase chain reaction (PCR), like any laboratory procedure, can be subject to a range of experimental or procedural error. A clear consideration of where such potential errors may occur is essential to minimize their impact. Careful quality control of equipment and reagents is essent ...

It is sometimes desirable to extract both RNA and DNA from the same sample, especially when the sample is small. This can be achieved by isolating a total nucleic acid fraction that is then divided into two portions, which are treated differentially with either Dnase I (to remove DNA and recover RNA) or with ...

There are two different methods of preparing tissue for histology: paraffin-embedding and freeze-embedding. Each has their advantages and drawbacks. Paraffin-embedded tissues (PET) produce optimum morphology but have comparatively poor molecular preservation and reco ...

Although individual microorganisms may well require a unique DNA extraction procedure, here we include robust techniques for the preparation of DNA from fungi, yeast, and bacteria, which yield DNA suitable for a PCR template.

The DNA extraction process represents one of the critical stages in the analysis of degraded or ancient DNA. If polymerase chain reaction (PCR) amplification starts from a poor extract containing low template quantities, stochastic variation in the amplification of individual alle ...

The successful extraction of viral RNA from biological material requires rapid transport and adequate storage of samples because of the unstable nature of RNA. Samples should be received and processed within 6 h and the relevant fractions stored at −70�C until testing. Also, it is difficult to a ...

In following any polymerase chain reaction (PCR)-based method, it is usual to identify the products of the reaction by some form of detection system. The majority of these still rely on size- and charge-based separation systems, although for some quantitative PCR applications, either direct ...

There are occasions where the only materiel available on a patient is stored plasma or serum samples. In normal individuals, the amount of DNA in these samples is very low but sufficient to serve as template for PCRs. Moreover, increased amounts of circulating DNA have been found in a variety of disorde ...

Although the best way of obtaining pure polymerase chain reaction (PCR) product will always be to optimize reaction conditions to yield only one product, there are still circumstances where DNA has to purifed from gels. Several good commercial products exist for the recovery of DNA from agaro ...

There are many differing protocols and a large number of commercially available kits used for the extraction of DNA from whole blood. This procedure is one we use routinely in both research and clinical service provision and is cheap and robust. It can also be applied to cell pellets from dispersed tis ...

The polymerase chain reaction (PCR) is a powerful method for fast in vitro enzymatic amplifications of specific DNA sequences. PCR amplifications can be grouped into three different categories: standard PCR, long PCR, and multiplex PCR. Standard PCR involves amplification of a single DNA ...

Based on the method of Chomczynski and Sacchi (1), this is an extremely reliable method without the requirement for centrifugation over CsCl gradients. As with any RNA protocol, extreme care should be taken to exclude RNAse contamination, the greatest source of which will be the sample itself. All ...