商品编号


L6030-W-L



品牌


Enzymatics



公司


Enzymatics,inc



公司分类


New Products



Size

24 Reactions

商品信息

WGS Ligase


Product Description

WGS Ligase is optimized for ligation following WGS Fragmentation. It catalyzes the formation of a phosphodiester bond between the terminal 5′ phosphate and a 3′ hydroxyl groups of duplex DNA or RNA. The enzyme efficiently joins blunt and cohesive ends and repairs single stranded nicks in duplex RNA, RNA, or DNA/RNA hydrid (1).

Source of Protein
A recombinant
E.coli
strain carrying the cloned T4 DNA Ligase gene

Unit Definition
1 unit is defined as the amount of DNA Ligase required to join 50% of 100 ng of DNA fragments with cohesive termini in 50 ?l 1X DNA Ligase Buffer following a 30 minute incubation at 23°C.

Molecular Weight
55,292 Daltons



Quality Control Analysis

Unit Activity
is measured using a 2-fold serial dilution method. Dilutions of enzyme batch were made in 1X DNA Ligase Reaction Buffer and added to 20 ?L reactions containing double stranded DNA fragments and 1X DNA Ligase Reaction Buffer. Reactions are incubated for 30 minutes at 23°C, stopped, and analyzed on a 1% agarose gel stained with ethidium bromide.

Protein Concentration
is determined by OD
280
absorbance.

Physical Purity
is evaluated by SDS-PAGE of concentrated and diluted enzyme solutions followed by silver stain detection. Purity is assessed by comparing the aggregate mass of contaminant bands in the concentrated sample to the mass of the protein of interest band in the diluted sample.

Single-Stranded Exonuclease
is determined in a 50 ?L reaction containing a r
ADI
olabeled single-stranded DNA substrate and 10 ?L of enzyme solution incubated for 4 hours at 37°C.

Double-Stranded Exonuclease
is determined in a 50 ?l reaction containing a r
ADI
olabeled double-stranded DNA substrate and 10 ?L of enzyme solution incubated for 4 hours at 37°C.

Double-Stranded Endonuclease
is determined in a 50 ?L reaction containing 0.5 ?g of plasmid DNA and 10 ?L of enzyme solution incubated for 4 hours at 37°C.

E.coli
16S rDNA Contamination
is evaluated using 5 ?L
r
eplicate samples of enzyme solution denatured and screened in a TaqMan qPCR assay for the presence of contaminating
E.coli
genomic DNA using oligonucleotide primers corresponding to the 16S rRNA locus.

Supplied in:
10 mM Tris-HCl
50 mM KCl
1 mM dithiothreitol
0.1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C

Supplied with:
B9020: 330mM Tris-HCl
50mM MgCl
5mM DTT
5mM ATP
30% PEG 6000
pH 7.6 @ 25°C


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Product Information



WGS Ligase


Part Number
L6030-W-L
Price
$261
Concentration
N/A
Unit Size
24 Reactions
Volume
0.24 mL


SDS

Available on request








Product Specification*


Storage Temperature
-25?C to -15?C


Test

Units Tested

Specification


SDS Purity
N/A
>99%
Specific Activity
N/A
300,000 U/mg
SS Exonuclease
6,000 U
< 1.0 % Released
DS Exonuclease
6,000 U
< 1.0 % Released
DS Endonuclease
6,000 U
No Conversion
E.coli
DNA Contamination
6,000 U
< 10 copies





* For a detailed summary of assay conditions and data, refer to the Quality Controls Analysis section.

References


Engler, M.J. and Richardson, C.C. (1982) P.D. Boyer (Eds.), The Enzymes, 5, pp. 3. San Diego: Academic Press.




Limitations of Use
This product was developed, manufactured, and sold for in vitro use only. The product is not suitable for admi
NIST
ration to humans or animals. SDS sheets relevant to this product are available upon request.