• 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)
  • Several plasmid vectors were developed to facilitate the expression and purification of such fusion proteins. (wikipedia.org)
  • Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms. (wikipedia.org)
  • The plasmid collection includes: Genome engineering CRISPRs Transcription Activator-Like Effector Nuclease (TALEN) kits Zinc finger nuclease kits Empty backbones Species-specific expression Epitope tags Fusion proteins Selectable markers Fluorescent marker Viral vectors Retroviral/Lentiviral Adenoviral AAV cDNA expression shRNA expression Molecular biology tools Vector Database-A curated list of over 4,000 vector backbones, including relevant cloning information and bacterial growth conditions. (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)
  • lysate
  • It is important that the lysate is clear at this stage to ensure good flow rates and, ultimately, to obtain protein-free plasmid DNA preparations. (qiagen.com)
  • 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)
  • 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)
  • capsid
  • Unlike viruses (which encase their genetic material in a protective protein coat called a capsid), plasmids are "naked" DNA and do not encode genes necessary to encase the genetic material for transfer to a new host. (wikipedia.org)
  • Molecular
  • The term plasmid was introduced in 1952 by the American molecular biologist Joshua Lederberg to refer to "any extrachromosomal hereditary determinant. (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)
  • A plasmid is a small DNA molecule within a cell that is physically separated from a chromosomal DNA and can replicate independently. (wikipedia.org)
  • 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)
  • viruses
  • Later in 1968, it was decided that the term plasmid should be adopted as the term for extrachromosomal genetic element, and to distinguish it from viruses, the definition was narrowed to genetic elements that exist exclusively or predominantly outside of the chromosome and can replicate autonomously. (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)
  • 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)
  • genes
  • While the chromosomes are big and contain all the essential genetic information for living under normal conditions, plasmids usually are very small and contain only additional genes that may be useful to the organism under certain situations or particular conditions. (wikipedia.org)
  • Many of the genes carried by a plasmid are beneficial for the host cells, for example: enabling the host cell to survive in an environment that would otherwise be lethal or restrictive for growth. (wikipedia.org)
  • The plasmid also carries genes to confer resistance to ampicillin and chloramphenicol. (wikipedia.org)
  • sequences
  • Sequence Analyzer-An Addgene software tool for creating plasmid maps from sequences with annotated features and restriction sites. (wikipedia.org)
  • identical
  • The size of the plasmid varies from 1 to over 200 kbp, and the number of identical plasmids in a single cell can range anywhere from one to thousands under some circumstances. (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)
  • Finally, the bound plasmid DNA is eluted in water or Tris-EDTA buffer. (sigmaaldrich.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)
  • genetic
  • This host-to-host transfer of genetic material is one mechanism of horizontal gene transfer, and plasmids are considered part of the mobilome. (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)
  • Miniprep of large number of plasmids can also be done conveniently on filter paper by lysing the cell and eluting the plasmid on to filter paper. (wikipedia.org)
  • Transformants retain the plasmid throughout cell divisions such that it can be recovered and used in other applications. (wikipedia.org)
  • Some plasmids, however, have no observable effect on the phenotype of the host cell or its benefit to the host cells cannot be determined, and these plasmids are called cryptic plasmids. (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
  • 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)