Applications
- Insertion of priming sites and conversion of single-copy clones to
multi-copy-inducible clones for higher yields of BAC or fosmid DNA.
The EZ-Tn5™ <oriV/KAN-2> Insertion Kit* randomly
inserts a single EZ-Tn5™ <oriV/KAN-2> Transposon containing
an inducible, high-copy origin of replication (oriV) into the
target DNA. Thus, the kit integrates CopyControl™ capability into
those vectors that lack it. TransforMax™ EPI300™ or EPI300™-T1R Electrocompetent E.
coli are transformed with an aliquot of the reaction and selected
for kanamycin resistance encoded by the EZ-Tn5 Transposon. Selected clones
are amplified by adding the CopyControl Induction or Autoinduction Solution,
and microgram quantities of template DNA can be purified from as little
as 1 ml of culture. |
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Figure 1 (click to enlarge). The process for generating EZ-Tn5™ <oriV/KAN-2> Transposon
insertion clones for high yields of DNA and bidirectional sequencing. |
