XcmI as a universal restriction enzyme for single-stranded DNA
Gene
|October 29, 1993
Abstract:
Single-stranded DNA can be cleaved into defined fragments at any predetermined site by interaction with a specially designed oligodeoxyribonucleotide (oligo) adaptor and the class-IIN restriction endonuclease, XcmI. The oligo adaptor has the structure [sequence: see text]. Upon hybridization to the target DNA through the central 9-nucleotide region and with the addition of XcmI, the template DNA is specifically cleaved to near completion. Hairpin structures on the template close to the hybridization site reduce the efficacy of cleavage.
Related Concept Videos
02:53Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
01:24DNA Isolation
DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
01:11Restriction Enzymes
Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...