ClaI

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Catalog Size Concentration
R0197L 5,000 units 5,000 units/ml
R0197S 1,000 units 5,000 units/ml

Recognition Site:

IMAGEM

Source:
An E. coli strain that carries the ClaI gene from Caryophanon latum L (ATCC 49862).

Reagents Supplied:
NEBuffer 4
BSA

Enzyme Properties
Activity in NEBuffers:
NEBuffer 1:        10%
NEBuffer 2:        50%
NEBuffer 3:        50%
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: Blocked by overlapping
dcm methylation: Not sensitive
CpG methylation: Blocked
More information about: Methylation Sensitivity

Heat Inactivation:
65°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 4
Supplemented with 100 μg/ml Bovine Serum Albumin
Incubate at 37°C.

1X NEBuffer 4:
20 mM Tris-acetate
50 mM potassium acetate
10 mM Magnesium Acetate
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 (dam-) in 1 hour at 37°C in a total reaction volume of 50 µl.

Concentration:
5,000 units/ml

Unit Assay Substrate:
λ DNA (dam-)

Storage Conditions:
10 mM Tris-HCl
50 mM KCl
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 A

Notes
General notes:

1. ClaI is an isoschizomer of BspDI.

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 ClaI, > 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 ClaI.

16-Hour Incubation:
A 50 μl reaction containing 1 μg of DNA and 200 units of ClaI 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 250 units of ClaI 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.

Endonuclease Activity:
Incubation of a 50 μl reaction containing 300 units of ClaI with 1 μg of ΦX174 RF I DNA for 4 hours at 37ºC resulted in < 0% conversion to RFII as determined by agarose gel electrophoresis.

Reagents Sold Separately
NEBuffer 4
BSA

Companion Products

dam-/dcm- Competent E. coli

Legal
Patents:
Invitrogen: Licensed Under U.S. Patent No. 5,312,746


ClaI FAQ

Q1: Is ClaI sensitive to methylation?
A1: Yes, it is blocked by overlapping dam methylation, and by CpG methylation at all sites.

Q2: Does ClaI produce commonly used compatible ends?
A2: BspDI cleaves to leave a 5´GC which can be ligated to fragments generated by AccI, BspDI, BstBI, TaqI, AciI, BsaHI, HinP1I, HpaII, NarI, Psp1406I.

Q3: Is ClaI an isoschizomer of BspDI?
A3: Yes, ClaI and BspDI recognize the same sequence, cut at the same site, both are blocked by dam methylation, use buffer 4, and are incubated at 37°. ClaI requires 5 units to cleave 1 μg pBR322 DNA versus 1 unit fron BspDI.

Q4: What is the activity of ClaI at 25°C?
A4: ClaI is 25-50% active at 25°C.

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