DNA purification kit

Repeat the phenol/chloroform/isoamylalcohol extraction step (Section 3.3.3 steps 3-5). Alternatively, use a DNA purification kit, which also provides good results. 

Plasmid is routinely recovered from bacteria by an alkaline lysis procedure, which lyses the bacterial cell while maintaining bacterial DNA attachment to the cell wall. This procedure enables subsequent precipitation of bacterial DNA and cellular debris, leaving a crude preparation enriched in plasmid. We routinely use a plasmid DNA purification kit provided by Qiagen that utilizes the alkaline lysis method for harvesting, and anion exchange column chromatography for rapid purification. We refer the reader to the detailed instructions provided in the kit by the manufacturer, which we have not found necessary to modify for purification of laboratory-use plasmid DNA. 

Affinity purify and desalt the DNA using the Qiagen QIAquick DNA Purification Kit. Add 1.0 mL of buffer QG (Qiagen) and mix. 

DNA purification kit [we used the Puregene Kit D-50KA (Centra, Minneapolis, MN) for flea and skin biopsy samples and the Qiaprep Spin Miniprep Kit (Qiagen) for the lymph node samples]. 

Harvest cells and extract DNA using a plasmid DNA purification kit (see Note 9). 

PCR PURIFICATION It is important to remove excess primers, nucleotides, enzymes, mineral oil, salts, and other impurities from DNA samples prior to executing other downstream processes. This DNA purification protocol is based on the QIAGEN QIAquick PCR Purification Kit (www.qiagen.com) however, several other DNA purification kits are available. We do not endorse this product but offer the outlined protocol as a starting place for the researcher. 

DNA Purification Kits QIAGEN s QIAquick 96-well PCR purification kit and QIAGEN s MinElute PCR purification kit. 

Separate aliquots of lOOg of pork-beef binary admixtures were prepared by mixing fresh pork and beef in a ratio of 100 0, 50 50, 25 75, 10 90, 5 95, 1 99, 0.5 99.5, 0.1 99.9 and 0 100 (w/w). The mixtures were autoclaved at 120°C for 2.5-h and DNAs were extracted from 100 mg sample in triplicates using MasterPure DNA Purification Kit (Epicentre Biotechnologies, Madison, USA) as per the protocol supplied by the manufacturer. The purity and concentration of extracted DNA samples were checked by Eppendorf UV-vis Biophotometer (Eppendorf, Germany). 

A prerequisite step to any rDNA work is the initial isolation of DNA or RNA from the source material (which can be microbial, plant, animal or viral). Numerous methodologies have been developed to achieve nucleic acid purification, and some of these methodologies have been adapted for use in a variety of commercially available purification kits. Although details vary, the general... 

PCR product purification kit (see 8.2.3.6 Cycle Sequencing of PCR-Amplified Genomic DNA with Universal Primers, subheading Specimen ). 

Polymerase chain reaction (PCR) purification kit, gel extraction kit, DNA mini-prep kit. 

To reduce false-positive clones, mRNA should be prepared free from rRNA and genomic DNA as possible. We purify and concentrate mRNA by QuickPrep Micro mRNA purification Kit (Pharmacia) using glycogen as a carrier. Then, contaminated gemonic DNA is digested with RNase-free DNase I (Gibco-BRL). 

QIAquick PCR Purification Kit (Qiagen) The features of the kit are available on the Web site http //www.qiagen.com/Products/ Catalog/Sample-Technologies/DNA-Sample-Technologies/ DNA-Cleanup/QIAquick-PCR-Purification-Kit. 

It is desirable to purify the PCR products prior to arraying in order to remove PCR-reaction components and cellular debris from the amplified target DNA, which would otherwise increase the printed spot size and interfere with the binding of the DNA to the slide surface. Several alternative methods could be used including (1) ethanol or isopropanol precipitation, or (2) 96-well PCR purification kits from TeleChem, Qiagen and Millipore. 

DNA isolation kits Wizard Plus Minipreps DNA Purification System (Promega, Madison, WI), Qiagen Plasmid Maxi kit (Qiagen, Chatsworth, CA). 

Amplified VH and VL DNA is purified using commercially available PureLink PCR purification kit and eluted in 100 pL volume. 

Pool all reactions and remove primers and other reaction components with commercially available PCR purification kit. Elute amplified DNA in lOOpLTE. SrrNote 11. 

DNA template preparations are a major source of contamination in cell-free synthesis. In vitro translation reactions are sensitive to contamination by salts, RNases, detergents, and alcohol, all of which are typically used in commercial plasmid purification kits. As such, plasmids prepared by resin-based purification protocols must be further purified by phe-nol/chloroform and chloroform extraction. DNA should be precipitated with isopropanol, washed in ethanol, dried, and taken up in RNase-free water to a concentration of about 1 mg mL . The DNA should be stored frozen, in small aliquots. 

Sensitivity Sensitivity has been determined by analyzing different types of matrices spiked with soy at decreasing levels of contamination (Table 17.5). On average, the limit of detection (LOD) resulted in 0.01 % (100 ppm) expressed as soy. In some cases it is possible to lower the limit to 0.005% (50 ppm), thanks to the high purification capability of DNA extraction kits. In this way, in fact, it is possible to add a higher... 

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