商品编号


TY0261H



品牌


Lucigen



公司


Lucigen



公司分类


In Vivo Tn5 Transposon Mutagenesis



Size

100 ?g

商品信息

Better transformation efficiencies with restriction system inhibition

Increase the insertion efficiency of EZ-Tn5? Transposome? complexes.

Enhance the transformation efficiency of unmodified DNA by blocking type I R-M systems, which are widespread in Eubacteria and Archaebacteria.




Applications

Increase the transformation efficiency of unmodified DNA
2
by blocking type I R-M systems, which are widespread in Eubacteria and Archaebacteria.
3

Increase the insertion efficiency of EZ-Tn5? Transposome? complexes.
2

DNA transformation can be difficult to achieve in many bacterial strains due to the presence of one or more restriction and modification (R-M) systems that cleave unmodified DNA that is recognized as "foreign." TypeOne? Restriction Inhibitor* provides a powerful method for increasing transformation efficiencies in bacterial strains with type I R-M systems. Developed as a unique preparation of a phage protein called ocr,
1
TypeOne Inhibitor is re
ADI
ly electroporated into cells along with transforming DNA.
In vivo
, the protein acts as a molecular decoy that blocks the DNA-binding site of type I R-M enzymes and inhibits DNA cleavage.

Benefits

Easy to use: Simply add TypeOne Restriction Inhibitor to unmodified DNA or a Transposome and electroporate using standard methods.

Blocks all known families of type I R-M enzymes
1
and can therefore be used to increase transformation efficiencies in a variety of bacterial cells.

Table 1. Effect of TypeOne? Restriction Inhibitor on transformation efficiencies.


Strain (Type I R-M system)


TypeOne?
Inhibitor

Type of DNA or Transposome?

Recombinants
per ?g DNA

S. typhimurium
LT2 (
Sty
L TIII)



pUC19 (100 pg)

3.0
×
10
6

S. typhimurium
LT2 (
Sty
L TIII)

X

pUC19 (100 pg)

3.0 × 10
8

E. coli
MG1655 (
Eco
K1)



48 kb fosmid (50 ng)

3.0
×
10
3

E. coli
MG1655 (
Eco
K1)

X

48 kb fosmid (50 ng)

1.4 × 10
6

S. typhimurium
LT2 (
Sty
L TIII)



EZ-Tn5? ori
/KAN-2>Tnp Transposome? (1 ?l)

1.3
×
10
4

S. typhimurium
LT2 (
Sty
L TIII)

X

EZ-Tn5? ori
/KAN-2>Tnp Transposome? (1 ?l)

1.0 × 10
6

References

Walkinshaw, M.D.
et al.
(2002)
Molec. Cell

9
, 187.

Hoff man, L.
et al.
(2002)
Epicentre Forum

9
(2), 8.

Murray, N.E.
et al.
(2000)
Microbial. Molec. Biol. Rev.

64
, 412.


*Covered by
issued and/or pending patents, exclusively licensed or assigned to Epicentre? (an
Illumina
? Company).