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Carboxy Flexi® Enzyme Blend (SgfI & EcoICRI)

R1901_Carboxy-Flexi--Enzyme-Blend-50-ul_3
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  • Available as a standalone product or as part of the HaloTag® Cloning Starter Kit (Cat.# G6050) or Carboxy Flexi® System, Transfer (Cat.# C9320)

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Catalog number selected: R1901

$ 123.00
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Carboxy Flexi® Enzyme Blend (SgfI & EcoICRI)
50μl
$ 123.00
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The Carboxy Flexi® Enzyme Blend (SgfI & EcoICRI) is required to cut acceptor Flexi® Vectors containing a carboxy-terminal peptide or protein tag. The donor vector (i.e., the clone containing the protein coding region of interest) is cut with SgfI and PmeI in a separate reaction. The two restriction digests are heat-inactivated and then combined with ligase to transfer the protein coding region to the acceptor vector, thereby appending the carboxy-terminal tag to the protein of interest. The ligated product no longer contains the EcoICRI site of the acceptor vector or the PmeI site of protein coding region, as these two blunt-end cutter target sites are not recreated in the ligation. Since the acceptor Flexi® Vector contains a lethal barnase gene, complete digestion with Carboxy Flexi® Enzyme Blend is not required for good transfer rates of the protein coding region from the donor Flexi® Vector.

SgfI recognition site:

GCG   ATCGC
CGCTA   GCG

EcoICRI recognition site:

GAGCTC
CTC GAG

5X Flexi® Digest Buffer: The composition of 5X Flexi® Digest Buffer is 50mM Tris-HCl (pH 7.9 at 37°C), 50mM MgCl2, 250mM NaCl, 5mM DTT, 0.5mg/ml acetylated BSA.

Storage Buffer: The Carboxy Flexi® Enzyme Blend (SgfI & EcoICRI) is supplied in 10mM Tris-HCl (pH 7.3 at 25°C), 150mM KCl, 25mM NaCl, 0.1mM EDTA, 1mM DTT, 0.5mg/ml BSA, 0.05% Thesit® and 50% glycerol.

Specifications

What's in the box?

Item Part # Size Available Separately

Nuclease-Free Water

P119A 1 × 1,250μl View Product

5X Flexi® Digest Buffer

R186A 1 × 1ml

Carboxy Flexi® Enzyme Blend (SgfI & EcoICRI)

R190A 1 × 50μl

Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Patents and Disclaimers

U.S. Pat. Nos. 8,293,503, 9,018,014 and 8,367,403, European Pat. No. 1685247 and other patents and patents pending.

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