HincII

Avalie o Produto
(0 votos)

 

Catalog Size Concentration
R0103L 5,000 units 10,000 units/ml
R0103S 1,000 units 10,000 units/ml

Recognition Site:

IMAGEM

Source:
A E. coli strain that carries the HincII gene from Haemophilus influenzae Rc (ATCC 49699).

Reagents Supplied:
NEBuffer 3 (10X)
BSA (100X)

Enzyme Properties
Activity in NEBuffers:
NEBuffer 1:        75%
NEBuffer 2:        100%
NEBuffer 3:        100%
NEBuffer 4:        100%

When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.

Methylation Sensitivity:
dam methylation: Not sensitive
dcm methylation: Not sensitive
CpG methylation: Blocked by some combinations of overlapping
More information about: Methylation Sensitivity

Heat Inactivation:
65°C for 20 minutes

Survival in a Reaction:
(+ + +) Suitable for an extended or overnight digestion. Enzyme is active > 8 hours.
More information about: Extended Digests with Restriction Enzymes

Reaction & Storage Conditions
Reaction Conditions:
1X NEBuffer 3
Supplemented with 100 μg/ml Bovine Serum Albumin
Incubate at 37°C.

1X NEBuffer 3:
50 mM Tris-HCl
100 mM NaCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.9 @ 25°C

Unit Definition:
One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Concentration:
10,000 units/ml

Unit Assay Substrate:
λ DNA

Storage Conditions:
10 mM Tris-HCl
200 mM NaCl
1 mM Dithiothreitol
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Storage Temperature:
-20°C

Diluent Compatibility:
Diluent B

Notes
General notes:

1. A 100-fold overdigestion of HincII in NEBuffer 4 produces star activity.

Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.

Ligation and Re-cutting:
After a 10-fold overdigestion with HincII, > 95% of the DNA fragments can be ligated with T4 DNA Ligase (at a 5' termini concentration of 1-2 μM) at 16ºC. Of these ligated fragments, > 95% can be recut with HincII.

16-Hour Incubation:
A 50 μl reaction containing 1 μg of DNA and 100 units of HincII incubated for 16 hours at 37ºC resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.

Exonuclease Activity:
Incubation of a 50 μl reaction containing 100 units of HincII with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (205 cpm/μg) for 4 hours at 37ºC released < 0.1% of the total radioactivity.

IMAGEM

HincII crystals (Ira Schildkraut and Lydia Dorner, New England Biolabs)

Reagents Sold Separately
NEBuffer 3
BSA

Legal
Patents:
New England Biolabs, Inc.: U.S. Patent No. 5,015,581
New England Biolabs, Inc.: U.S. Patent No. Re. 35248


HincII FAQ

Q1: Do degenerate recognition sites need to be palindromic?
A1: Most restriction enzyme recognition sites are palindromic and include only specified base pairs (i.e., EcoRI recognizes GAATTC). However, some enzymes have degenerate sites, meaning that they contain one or more base pairs that are not specifically defined (i.e., BsrFI recognizes RCCGGY, where R= A or G and Y= C or T). For degenerate enzymes, any base represented by the single letter code may be present at either location in the recognition site for cleavage to occur. For example, BsrFI recognizes all of the following sequences: ACCGGC, ACCGGT, GCCGGC, GCCGGT.

 

DESENHE SEUS PRIMERS!

Pesquise seu item

 

Receba Promoções

Assistência Técnica