商品编号


C300C105



品牌


Lucigen



公司


Lucigen



公司分类


CRISPR, Large or Difficult Fragment Cloning




10 X 50 ?l



商品信息

Standard cloning for large inserts and Gibson assembly electroporation

Clone large inserts and transform large plasmids - up to at least 145 kb plasmid DNA.

Generate clones with induc
IBL
e copy number using CopyControl? vectors.

Achieve high transformation efficiencies: >1 × 10
10
cfu/?g pUC19 DNA.

Increase Gibson Assembly? colony counts with high-efficiency electroporation





Free Sample Available




Applications

Generation of induc
IBL
e copy-number clones using the CopyControl? Cloning System.


TransforMax? EPI300?
E. coli
cells lack the
tonA
gene and are engineered for use with Epicentre's CopyControl?
CDN
A, Gene, and PCR Cloning Kit and other CopyControl Cloning Systems* that do not require phage T1-resistant cells. The cells contain an induc
IBL
e mutant
trfA
gene whose gene product is required for initiation of replication from the
ori
V origin of replication, such as that in CopyControl pCC1? vectors. On LB chloramphenicol plates or in LB or SOC media supplemented with chloramphenicol, CopyControl clones grown in TransforMax EPI300
E. coli
replicate at single-copy number from the F-factor replicon because expression of the
trfA
gene is repressed. Addition of CopyControl Induction Solution induces the cells to express the
trfA
gene product and induces their replication at high-copy number from
ori
V (Fig. 1). Replication from
ori
V results in higher yields and higher purity of cloned DNA.

Benefits

Copy-number of clones is under tight control of an induc
IBL
e promoter linked to the
trfA
gene.

High transformation efficiency with clones of all sizes.

lacZΔM15
for blue/white screening of recombinants.

Restriction-minus [
mcrA, Δ(mrr-hsdRMS-mcrBC
)] phenotype enables efficient cloning of methylated DNA.

Endonuclease-minus (
endA1
), to ensure high yields of DNA.

Recombination-minus (
recA1
), for greater st
ABI
lity of large insert clones.





Figure 1. Copy number of CopyControl? BAC clones can be induced 10- to 20-fold in TransforMax? EPI300?
E. coli.
The yield of BAC DNA from CopyControl BAC clones of 110-145 kb increased >14-fold following addition of CopyControl Induction Solution. U=uninduced cells; I=induced cells.

Genotype

F
-
mcrA Δ(mrr-hsdRMS-mcrBC) Φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ
-
rpsL (Str
R
) nupG trfA dhfr

TransforMax EPI300 Electrocompetent
E. coli

Transformation efficiency of >1 x 10
10
cfu/?g of pUC19.

TransforMax EPI300 Chemically Competent
E. coli

Transformation efficiency of >5 x 10
8
cfu/?g of pUC19.

*Covered by issued and/or pending patents
.


ORDER INFORMATION
1,000X concentrated solution. Filter sterilized.