Notes: Sputum samples were collected from cystic fibrosis patients and 16S rRNA sequences amplified by PCR. These products were cloned into a T-vector, transformed into competent cells and the resulting colonies grown in 2ml LB broth in 96-deep-well plate for 20 hours. Of this culture, 1.9ml was pelleted and the clones isolated using the Wizard® SV Plasmid Purification System. The purified plasmid DNA was subjected to agarose gel electrophoresis and sequenced. (4133)

Notes: The authors use the Flexi^® Cloning System to convert their cDNA clones to expression-ready clones. They wanted clones that could be used for comprehensive analysis with the HaloTag^® Technology. They also describe a method of transferring ORFs between Flexi^® Vectors in a 96-well plate format. They also used Wizard^® SV 96 Plasmid DNA Purification, Wizard^® SV PCR Clean-Up, and Wizard^® SV Gel and PCR Clean-Up Systems. (4056)

Notes: The authors examined the pattern of Anaplasma phagocytophilum msp2 expression, a gene that modulates with little immune pressure and has decreased virulence with prolonged in vitro passage. C57BL/6J mice were inoculated with HL-60 cells infected with low-passage (passage 5) or high-passage (passage 26). Blood samples were taken 2–21 days post-inoculation, and total RNA was isolated. The purified RNA was subjected to RT-PCR, cloned into the pGEM^®-T Easy Vector, transformed and plated. Plasmids were purified using the Wizard^® SV 96 Plasmid DNA Purification System, and the insert size analyzed after EcoRI digestion. The inserts were sequenced, aligned with A. phagocytophilum Webster strain msp2 references using ClustalX and the diversity of msp2 transcripts divided into low- or high-passage bacteria. (3975)

Notes: This study compared detection of Pythium species in soil samples by DNA array hybridization and PCR cloning. Three Pythium species were amplified from soil samples, a single 3´ A was added to the resulting PCR product, and the DNA was ligated into the pGEM^®-T Easy Vector at 4°C overnight. After the ligation was transformed into JM109 Competent Cells, and 100 colonies were chosen and grown overnight in LB broth. The plasmid DNA was isolated using the Wizard^® SV 96 Plasmid DNA Purification System and then sequenced. (3437)

Notes: The Wizard^® SV 96 Plasmid Purification System was used to purify plasmid DNA on the Beckman Coulter Biomek^® NX Laboratory Automation Workstation. Plasmid DNA quantity and quality were assessed by spectrophotometry, restriction digestion and PCR. The Wizard^® SV 96 Plasmid DNA Purification System was also used to purify plasmid DNA from bacterial clones isolated in a bacterial two-hybrid screening. (3300)

Notes: To generate mini-arrays, the Wizard^® SV 96 Plasmid DNA Purification System was used to isolate BAC DNAs from bacterial clones. The authors implemented the Wizard^® system using a Biomek 2000 Laboratory Automation workstation, and comment that the resulting DNA had low levels of contamination from the host E. coli, and was suitable for DNA amplification and subsequent array production. The performance of the mini-arrays was evaluated for gains and losses, with emphasis on reducing assay time and the amount of target DNA. They found that gains and losses of relatively large genomic regions such as full chromosomes and chromosomal arms could be reliably detected in <4 hours. (3542)

Notes: The Wizard^® SV 96 Plasmid DNA Purification System was used to purify cosmid DNA from JM109 transformed bacteria. Five hundred nanograms of each purified cosmid were used in sequencing reactions. (3207)

Notes: The Wizard^® SV 96 Plasmid DNA Purification System was used to purify random subtractive PCR clones from DH5α cells.  The subtractive PCR clones were made from porcine spherical and tubular conceptuses.  Purified clones were denatured and screened with a DIG High Prime DNA Labeling and Detection Starter Kit (Roche Applied Science). (2748)

Notes: Researchers used the Wizard^® SV 96 Plasmid Purification System to purify plasmids carrying BAC clone sequences from rice and maize positively identified by a “gene-island” sequencing strategy. Two microliters of the purified plasmids were added to sequencing reactions. Sequencing was performed using the ABI BigDye^® terminator mix and an ABI 9700 thermal cycler. Sequencing reaction products were separated on an ABI 3700 DNA sequencer. (2677)