Are BamHI and BglII compatible?
The end of a DNA fragment cut with BamHI and BglII (black arrows) has a 50% chance of making a BamHI-BglII junction and a 25% chance of making either identical-end junction. Self-ligation will produce a circular molecule.
What are compatible restriction enzymes?
As unlikely as it may seem, restriction enzymes from different organisms can produce interlocking pieces of DNA – so called compatible cohesive ends (CCE). These are pieces of DNA, which fit together and can be ligated, creating a hybrid molecule.
What restriction enzymes are compatible ends?
Compatible Cohesive Ends and Generation of New Restriction Sites
| Enzyme | Ligated To | Recleaved By |
|---|---|---|
| BglII (A/GATCT) | BclI, DpnII | DpnII |
| BsaHI | ||
| (GR/CGYC) | AccI (GT/CGAC), ClaI, BstBI, TaqI-v2 | — |
| (GA/CGYC) | AciI, HinP1I | HgaI |
Does the restriction enzyme BamHI produce sticky or blunt ends?
Recognition Sequences
| Enzyme | Organism | Blunt or Sticky End |
|---|---|---|
| EcoRI | Escherichia Coli | Sticky |
| BamHI | Bacillus amyloliquefaciens | Sticky |
| BglII | Bacillus globigii | Sticky |
| PvuI | Proteus vulgaris | Sticky |
What is the difference between blunt ends and sticky ends of DNA cut by restriction enzymes?
Answer: Blunt and sticky ends areresult of restriction endonuclease action on double stranded DNA. Sticky Ends – are staggered ends on a DNA molecule with short, single-stranded overhangs. Blunt Ends are a straight cut, down through the DNA that results in a flat pair of bases on the ends of the DNA.
Why sticky end is better than blunt end?
The advantage of sticky ends is that a fragment of human DNA can only fit into a bacterial plasmid in one direction. In contrast, if both the human DNA and bacterial plasmid have blunt ends, the human DNA can be inserted head-to-tail or tail-to-head into the plasmid.
What are the reaction conditions for xmaji ( avrii )?
Thermo Scientific XmaJI (AvrII) restriction enzyme recognizes C^CTAGG sites and cuts best at 37°C in Tango buffer. See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes.
Which is the high fidelity version of nhei?
NheI has a High Fidelity version NheI-HF® ( NEB #R3131 ). High Fidelity (HF) Restriction Enzymes have 100% activity in CutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing.
How is CTAG cleaved in avrii and XbaI?
Cleaves to leave a 5´ CTAG extension which can be efficiently ligated to DNA fragments generated by AvrII, SpeI or XbaI. Activity inhibited by salt concentrations > 100 mM.
Which is E coli strain carries the avrii gene?
Product Source An E. coli strain that carries the AvrII gene from Anabaena variabilis UW (E. Rosenvold). Properties & Usage Unit Definition One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA (HindIII digest) in 1 hour at 37°C in a total reaction volume of 50 µl.
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