• vectors
  • Virtually all plasmid vectors in common use encode one or more antibiotic resistance genes as a selectable marker, for example a gene encoding ampicillin or kanamycin resistance, which allows bacteria that have been successfully transformed to multiply uninhibited. (wikipedia.org)
  • Vectors are generally derived from plasmids or viruses, and represent relatively small segments of DNA that contain necessary genetic signals for replication, as well as additional elements for convenience in inserting foreign DNA, identifying cells that contain recombinant DNA, and, where appropriate, expressing the foreign DNA. (wikipedia.org)
  • Digestion
  • Plasmid DNA is ready for immediate use in restriction enzyme digestion (Figure 1) cloning, PCR, transcription, conventional and automated sequencing (Figure 2) . (sigmaaldrich.com)
  • This denatured form of the plasmid runs faster on agarose gels and is resistant to restriction enzyme digestion. (qiagen.com)
  • Plasmid Cloning Guides Molecular Cloning Guides-References to help scientists design plasmid cloning experiments, including tutorials on restriction enzyme digestion and PCR-based cloning. (wikipedia.org)
  • bacteria
  • Bacteria that have not taken up the plasmid vector are assumed to lack the resistance gene, and thus only colonies representing successful transformations are expected to grow. (wikipedia.org)
  • In a process called transformation, plasmid DNA is taken up by the bacteria under a variety of conditions. (wikipedia.org)
  • Without functional RecA protein, the exogenous plasmid DNA is left unaltered by the bacteria. (wikipedia.org)
  • vector
  • Once the vector is set up with flanking restriction sites, all manipulations (i.e., mutagenesis, sequencing, expression) can be performed in the same plasmid. (wikipedia.org)
  • chromosomal
  • The optimized lysis time allows maximum release of plasmid DNA from the cell without release of cell wall-bound chromosomal DNA, while minimizing the exposure of the plasmid to denaturing conditions. (qiagen.com)
  • Separation of plasmid from chromosomal DNA is based on coprecipitation of the cell wall-bound chromosomal DNA with the insoluble complexes containing salt, detergent, and protein. (qiagen.com)
  • Since chromosomal fragments are chemically indistinguishable from plasmid DNA under the conditions used, the two species will not be separated on QIAGEN resin and will elute under the same salt conditions. (qiagen.com)
  • Genome
  • Its plasmid collection contains plasmids used for functions such as genome engineering (including CRISPRS), gene expression, shRNA knockdown, viral-mediated gene delivery, detection of miRNA and promoter activity. (wikipedia.org)
  • facilitates
  • Addgene facilitates the exchange of genetic material between laboratories by offering plasmids and their associated cloning data to not-for-profit laboratories around the world. (wikipedia.org)
  • buffers
  • NucleoBond Xtra columns: The enlarged columns lead to lower silica resin beds and enable faster flow of lysate and buffers through the columns, allowing for a fast purification. (clontech.com)
  • coli
  • The NucleoSpin 96 Plasmid Kits are designed for purifying up to 20 µg of high-copy plasmids from 1-5 ml E. coli overnight culture, preferably grown in LB medium. (clontech.com)
  • E. coli strains are often genetically modified to contain a mutant recA allele and thereby ensure the stability of extrachromosomal segments of DNA, known as plasmids. (wikipedia.org)
  • Plasmid pHT01 is generally stable in both B. subtilis and Escherichia coli, and can be used for protein expression in these host strains. (wikipedia.org)
  • restriction
  • Sequence Analyzer-An Addgene software tool for creating plasmid maps from sequences with annotated features and restriction sites. (wikipedia.org)
  • buffer
  • After washing, the purified plasmid DNA is eluted in a high-salt buffer and precipitated with isopropanol. (clontech.com)
  • The plasmid DNA is then efficiently eluted from the QIAGEN-tip with high salt buffer (Buffer QF or QN). (qiagen.com)
  • Highly pure plasmid DNA is eluted with Elution Buffer AE (5 mM Tris / HCl, pH 8.5) or water (pH 8.0- 8.5), and can directly be used for downstream applications. (clontech.com)
  • biomolecules
  • Depending on the column size, CIM monolithic columns are primarily used for the purification or analysis of large biomolecules which are being used for cancer gene therapy, treatment of viral infectious diseases and treatment of genetic diseases. (wikipedia.org)
  • cell
  • I am purifying plasmid DNA using Promega miniprepa DNA Purification System from Novablue cell transformated by Clontech pEGFP-N1 containing the insert. (bio.net)
  • Transformants retain the plasmid throughout cell divisions such that it can be recovered and used in other applications. (wikipedia.org)
  • The MS2-GFP fusion protein was expressed by transferring it to a cell with a plasmid (Robert Singer's lab). (wikipedia.org)
  • scientists
  • The organization covers the operating costs of maintaining and improving the collection by charging a nominal fee to scientists requesting plasmids. (wikipedia.org)
  • process
  • In this way, the purification time can be significantly decreased, resulting in a pronounced reduction of the purification process costs. (wikipedia.org)
  • By using the standard Universal Biological Material Transfer Agreement (UBMTA) and implementing electronic signatures, Addgene's electronic MTA (eMTA) system expedites the approval process for plasmid orders. (wikipedia.org)