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  • enzymes
  • The DNA fragment to be mutated is inserted into a phagemid such as M13mp18/19 and is then transformed into an E. coli strain deficient in two enzymes, dUTPase (dut) and uracil deglycosidase (ung). (wikipedia.org)
  • Both enzymes are part of a DNA repair pathway that protects the bacterial chromosome from mutations by the spontaneous deamination of dCTP to dUTP. (wikipedia.org)
  • In RFLP analysis, the DNA sample is broken into pieces (and digested) by restriction enzymes and the resulting restriction fragments are separated according to their lengths by gel electrophoresis. (wikipedia.org)
  • Nobel prize winner Arthur Kornberg used ΦX174 as a model to first prove that DNA synthesized in a test tube by purified enzymes could produce all the features of a natural virus, ushering in the age of synthetic biology. (wikipedia.org)
  • gene
  • Site-directed mutagenesis is a molecular biology method that is used to make specific and intentional changes to the DNA sequence of a gene and any gene products. (wikipedia.org)
  • The single-strand primer is then extended using a DNA polymerase, which copies the rest of the gene. (wikipedia.org)
  • When scientists transfer a gene from one cell into another cell in order to express the new genetic material as a protein in the recipient cell, the cDNA will be added to the recipient (rather than the entire gene), because the DNA for an entire gene may include DNA that does not code for the protein or that interrupts the coding sequence of the protein (e.g., introns). (wikipedia.org)
  • If researchers were trying to initially determine the chromosomal location of a particular disease gene, they would analyze the DNA of members of a family afflicted by the disease, and look for RFLP alleles that show a similar pattern of inheritance as that of the disease (see Genetic linkage). (wikipedia.org)
  • genome
  • In the first schematic, a small segment of the genome is being detected by a DNA probe (thicker line). (wikipedia.org)
  • The phi X 174 (or ΦX174) bacteriophage is a single-stranded DNA (ssDNA) virus and the first DNA-based genome to be sequenced. (wikipedia.org)
  • As it translocates around the genome it displaces the outer strand of already-synthesised DNA, which is immediately coated by SSBP proteins. (wikipedia.org)
  • Subsequently, the generated DNA is translocated into the nucleus of the host cell where it is integrated in its genome by the retroviral integrase. (wikipedia.org)
  • degenerate
  • Would I need to use a degenerate/wobble base substitution to take this possibility into account ie: Y sub for C/T or would the single base mismatch in the middle of the sequence still allow the primer to bind? (protocol-online.org)
  • assay
  • Pools of pre-designed primers can help generate comprehensive expression profiles, especially where assay sensitivity is critically important. (thermofisher.com)
  • The present invention provides these E. suis primers and a method to use these primers in a PCR protocol to provide a highly sensitive diagnostic assay for early signs of an E. suis infection. (google.com)
  • The toeprinting assay, also known as the primer extension inhibition assay, is a method used in molecular biology that allows you to examine the interactions between messenger RNA and ribosomes or RNA-binding proteins. (wikipedia.org)
  • It is different from the more commonly used DNA footprinting assay. (wikipedia.org)
  • A schematic of a toeprinting assay can be found here: https://www.researchgate.net/figure/41124064_fig2_A-Schematic-of-the-primer-extension-inhibition-toeprint-assay-illustrating-how. (wikipedia.org)
  • cDNA
  • The Supreme Court also noted that "cDNA is not a 'product of nature' and is patent eligible under §101, except insofar as very short series of DNA may have no intervening introns to remove when creating cDNA. (biologicsblog.com)
  • According to the Court, "[i]n that situation, a short strand of cDNA may be indistinguishable from natural DNA. (biologicsblog.com)
  • The term cDNA is also used, typically in a bioinformatics context, to refer to an mRNA transcript's sequence, expressed as DNA bases (GCAT) rather than RNA bases (GCAU). (wikipedia.org)
  • An example of this first step from viral DNA to cDNA can be seen in the HIV cycle of infection. (wikipedia.org)
  • fragment
  • A dark band in a lane indicates a DNA fragment that is the result of chain termination after incorporation of a dideoxynucleotide (ddATP, ddGTP, ddCTP, or ddTTP). (wikipedia.org)
  • Its limitations include dye effects due to differences in the incorporation of the dye-labelled chain terminators into the DNA fragment, resulting in unequal peak heights and shapes in the electronic DNA sequence trace chromatogram after capillary electrophoresis (see figure to the left). (wikipedia.org)
  • probe
  • The program is a comprehensive real time PCR primer and probe search and analysis tool, and also does other tasks such as siRNA and molecular beacon searches, open reading frame and restriction enzyme analysis etc. (wikipedia.org)
  • heteroduplex
  • The heteroduplex DNA, that forms, consists of one parental non-mutated strand containing dUTP and a mutated strand containing dTTP. (wikipedia.org)
  • The other role of the RT is its ribonuclease H activity that degrades RNA only when it is in a heteroduplex with DNA. (wikipedia.org)
  • Mesh
  • DNA Primers" is a descriptor in the National Library of Medicine's controlled vocabulary thesaurus, MeSH (Medical Subject Headings) . (ucdenver.edu)
  • Detection
  • Detection of DNA in specimens comprising body fluids of tissues can be difficult because of the small quantity of DNA present or because of the presence in the specimen of other interfering materials, including DNA from a different source. (google.com)
  • The basic technique for the detection of RFLPs involves fragmenting a sample of DNA by a restriction enzyme, which can recognize and cut DNA wherever a specific short sequence occurs, in a process known as a restriction digest. (wikipedia.org)
  • Irrespective of the variety of methods used for DNA analysis, only PCR in its different formats has been widely applied in GMO detection/analysis and generally accepted for regulatory compliance purposes. (wikipedia.org)
  • An event-specific detection searches for the presence of a DNA sequence unique to a certain GMO, usually the junction between the transgene and the organism's original DNA. (wikipedia.org)
  • The construct-specific detection methods can either be DNA or protein based. (wikipedia.org)
  • DNA based detection looks for a part of the foreign DNA inserted in a GMO. (wikipedia.org)
  • Currently, it is highly unlikely that the presence of unexpected or even unknown GMOs will be detected, since either the DNA sequence of the transgene or its product, the protein, must be known for detection. (wikipedia.org)
  • commonly
  • DNA profiling is most commonly used as a forensic technique in criminal investigations to identify an unidentified person or whose identity needs to be confirmed, or to place a person at a crime scene or to eliminate a person from consideration. (wikipedia.org)
  • Q-PCR is commonly used to determine whether a DNA sequence is present in a sample and the number of its copies in the sample. (wikipedia.org)
  • genomic
  • Myriad sued Ambry shortly after the Supreme Court held in Association for Molecular Pathology v. Myriad (Myriad) that Myriad's claims to cDNAs were patent eligible as long as the cDNAs did not have the same sequence as natural DNAs, but that Myriad's claims to isolated genomic DNA (DNA with the same sequence as natural DNA) were not patentable. (biologicsblog.com)
  • single-strand
  • As the double-mutant E. coli replicates the phage DNA, its enzymatic machinery may, therefore, misincorporate dUTP instead of dTTP, resulting in single-strand DNA that contains some uracils (ssUDNA). (wikipedia.org)
  • By convention, if the base sequence of a single strand of DNA is given, the left end of the sequence is the 5' end, while the right end of the sequence is the 3' end. (wikipedia.org)
  • Genetics
  • The modern process of DNA profiling was developed in 1984 by Sir Alec Jeffreys while working in the Department of Genetics at the University of Leicester. (wikipedia.org)
  • The first methods for finding out genetics used for DNA profiling involved RFLP analysis. (wikipedia.org)
  • synthesize
  • RNA primers could be used but they are unstable and Rnases are frequent and really hard to get rid of, while DNA is easy to synthesize artificially and is way more stable. (biology-online.org)
  • To synthesize an additional DNA strand, traditionally one would digest the RNA of the hybrid strand, using an enzyme like RNase H, or through alkali digestion method. (wikipedia.org)
  • enzyme
  • DNA is collected from cells, such as a blood sample, and cut into small pieces using a restriction enzyme (a restriction digest). (wikipedia.org)
  • Electrophoresis
  • Annealing temperatures for each of the primer sets must be optimized to work correctly within a single reaction, and amplicon sizes, i.e., their base pair length, should be different enough to form distinct bands when visualized by gel electrophoresis. (wikipedia.org)
  • protein
  • A Protein Sequencer using Edman degradation was installed in 1989 quickly followed by several DNA Sequencers which were the first to use fluorescent dye terminator chemistry. (wikipedia.org)
  • molecular
  • Primers are used extensively in genetic and molecular biology techniques. (mcw.edu)
  • Sarah H. Haddock, Christine Quartararo, Patrick Cooley, and Dat D. Dao (2002) Low-Resolution Typing of HLA-DQA1 Using DNA Microarray, Methods in Molecular Biology 170, 201-210. (wikipedia.org)
  • RFLP
  • RFLP tests require much bigger samples of DNA than do short tandem repeat (STR) tests. (wikipedia.org)
  • reactions
  • This is frequently performed using a denaturing polyacrylamide-urea gel with each of the four reactions run in one of four individual lanes (lanes A, T, G, C). The DNA bands may then be visualized by autoradiography or UV light and the DNA sequence can be directly read off the X-ray film or gel image. (wikipedia.org)
  • specific
  • Developed by Kary Mullis in 1983, a process was reported by which specific portions of the sample DNA can be amplified almost indefinitely (Saiki et al. (wikipedia.org)
  • PCR greatly amplifies the amounts of a specific region of DNA. (wikipedia.org)
  • Limitations include non-specific binding of the primer to the DNA, affecting accurate read-out of the DNA sequence, and DNA secondary structures affecting the fidelity of the sequence. (wikipedia.org)
  • dNTPs
  • In their respective triphosphate forms, NRTIs and the only NtRTI available compete with their corresponding endogenous deoxynucleotide triphosphate (dNTPs) for incorporation into the nascent DNA chain (see figure 1). (wikipedia.org)