BtgI

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Catalog Size Concentration
R0608L 5,000 units 10,000 units/ml
R0608S 1,000 units 10,000 units/ml

Recognition Site:

IMAGEM

Source:
An E. coli strain that carries the cloned BtgI gene from Bacillus thermoglucosidasius (X. Pan)

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

Enzyme Properties
Activity in NEBuffers:
NEBuffer 1:        25%
NEBuffer 2:        50%
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: Not sensitive
More information about: Methylation Sensitivity

Heat Inactivation:
80°C for 20 minutes

Survival in a Reaction:
(+) Intermediate activity. Suitable for extended digestion, but < 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 pBR32 DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Concentration:
10,000 units/ml

Unit Assay Substrate:
pBR322 DNA

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

Storage Temperature:
-20°C

Diluent Compatibility:
Diluent B

Notes
General notes:

1. DsaI is an isoschizomer of BtgI.

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 60-fold overdigestion with BtgI, > 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 BtgI.

16-Hour Incubation:
A 50 μl reaction containing 1 μg of DNA and 64 units of BtgI 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 125 units of BtgI 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.

Reagents Sold Separately
NEBuffer 3
BSA


BtgI 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.

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