hybridization

Examples of hybridization in the following topics:

• Two-Hybrid Analysis

  • The two-hybrid method detects the interaction of two proteins by their ability to reconstitute the activity of a split transcription factor.
  • The most widely employed tools are the yeast two-hybrid system .
  • The yeast two-hybrid screening system is an effective and quick tool for the in vivo study of protein–protein interaction both in prokaryotes and eukaryotes.
  • One limitation of classic yeast two-hybrid screens is that they are limited to soluble proteins.
  • Overview of two-hybrid assay, checking for interactions between two proteins, called here Bait and Prey.

• The FISH Technique

  • FISH is a hybridization technology which allows the labeling of target RNAs with a fluorescent probe.
  • FISH (fluorescence in situ hybridization) is a cytogenetic technique developed by biomedical researchers in the early 1980s.
  • The probe must be large enough to hybridize specifically with its target but not so large as to impede the hybridization process.
  • They are anti-sense to the target mRNA or DNA of interest, thus they hybridize to targets.
  • A similar hybridization technique is called a zoo blot.

• Whole-Genome DNA-Binding Analysis

  • Two different fluorescence labels are used to label the IP DNA, and a hybridization-control DNA, respectively.
  • Usually, total DNA before IP (input DNA) is used as hybridization control.
  • The two differentially-labeled DNAs are hybridized to the same microarray and the difference in fluorescence intensity gives a measure of the enrichment .

• DNA Sequencing of Insertion Sites

  • These include Southern hybridization, inverse Polymerase Chain Reaction (iPCR), and most recently, vectorette PCR to identify and map the genomic positions of the insertion sequences.
  • Southern hybridization is rather time-consuming and requires additional procedures for localizing ISs.
  • Also, the length of each restriction DNA fragment containing a target sequence must be determined by Southern hybridization followed by sub-genomic fractioning before intramolecular ligation and PCR amplification.
  • These difficulties render Southern hybridization and iPCR impractical as techniques for quickly surveying repetitive elements in genomes.

• Mapping Protein-Protein Interactions

  • The most widely employed tools are the yeast two-hybrid system and affinity purification coupled to mass spectrometry.
  • The yeast two-hybrid screening system is an effective and quick tool for the in vivo study of protein–protein interaction both in prokaryotes and eukaryotes.

• Allergic Responses to Drugs

  • In the process of sp3 hybridization in methane, the single 2s and three 2p orbitals of carbon mix into four sp3 hybrid orbitals, which are chemically and geometrically identical.

• DNA Analysis Using Genetic Probes and PCR

  • Hybridization of the sequence with a complementary sequence of DNA or RNA, follows cleavage of the double-stranded DNA of the microorganism in the specimen.
  • In many PCR-based typing assays, the target DNA of interest is amplified and labeled by PCR, and the labeled products are hybridized to an array of immobilized diagnostic probes.

• Genetic Analysis

  • Microarray hybridization is another technique used to characterize the dynamic nature of gene expression within a microbial cell.
  • Summarize the techniques used to study genomes: PFGE. ordered clone approach, direct shotgun sequencing and microarray hybridization

• Northern Blots

  • Northern blotting involves the use of electrophoresis to separate RNA samples by size and detection with a hybridization probe complementary to part of or the entire target sequence.

• Reporter Fusions

  • A reporter fusion is the hybrid of a gene or portion of a gene with a tractable marker.
  • The resulting protein-GFP hybrid transcribed from the reporter construct now has a protein attached to GFP.