Description
Sample material:≤200 ng sticky-ended or blunt-ended DNA
Incubation temperature: +15°C to +25°C
Incubation time: 5 min
Product: Ligated DNA, suitable for direct use in transformation experiments
Specifications/Features
Information:
All necessary reagents are required for ligation are provided. There is no need to prepare buffers or to add ATP or Mg2+
The efficiency of the Rapid DNA Ligation Kit from
Roche Molecular Biochemicals was compared with
standard and optimized conventional ligation procedures.
The reactions generated circular recombinant
bacteriophage DNA in a three-fragment assembly
DNA ligation. Sequential blunt-end and cohesive-end
ligation, including a temperature shift from 26°C to
14°C, led to a more than 10-fold increase in the number
of transformants obtained. Applying a modified
protocol, the kit was found to provide a reliable, fast,
and remarkably efficient procedure for this complex
application.
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Simplify dephosphorylation of 5′ ends on DNA vectors by
using our high-quality Shrimp Alkaline Phosphatase.
Avoid phenol:chloroform extractions; after dephosphorylation,
inactivate Shrimp Alkaline Phosphatase completely
and irreversibly by heating at 65 °C for only 15 minutes.
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For most cloning procedures, ligation reactions are conventionally performed at 16ºC overnight; this...
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I first tried the Roche/Boehringer Rapid Ligation Kit in the first year of my Ph.D., over 4 years ...
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