|R0197L||5,000 units||5,000 units/ml|
|R0197S||1,000 units||5,000 units/ml|
An E. coli strain that carries the ClaI gene from Caryophanon latum L (ATCC 49862).
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.
dam methylation: Blocked by overlapping
dcm methylation: Not sensitive
CpG methylation: Blocked
More information about: Methylation Sensitivity
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
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
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.
Unit Assay Substrate:
λ DNA (dam-)
10 mM Tris-HCl
50 mM KCl
1 mM Dithiothreitol
0.1 mM EDTA
200 µg/ml BSA
pH 7.4 @ 25°C
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.
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.
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.
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
dam-/dcm- Competent E. coli
Invitrogen: Licensed Under U.S. Patent No. 5,312,746
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.