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Restriction Endonucleases >
Restriction Endonucleases >
PmlI |
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Prices are in US dollars and valid only for US orders.
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Recognition Site:
isoschizomers | compatible ends | single letter code
Source:
Pseudomonas maltophilia (T. Tenkanen)
Reagents Supplied:
NEBuffer 1
BSA
Enzyme Properties
Activity in NEBuffers:
| NEBuffer 1: | | 100% | | NEBuffer 2: | | 75% | | NEBuffer 3: | | 0% | | NEBuffer 4: | | 75% |
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 |
Heat Inactivation:
65°C for 20 minutes
Survival in a Reaction:
Minimum units to digest 1 µg of substrate DNA in 16 hours: 0.50 unit(s)
Reaction & Storage Conditions
Reaction Conditions:
1X NEBuffer 1
Supplemented with 100 μg/ml Bovine Serum Albumin
Incubate at
37°C.
1X NEBuffer 1:
10 mM Bis-Tris-Propane-HCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.0 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme required to digest 1 µg
l DNA (HindIII digest) DNA in 1 hour at 37°C in a total reaction volume of 50 µl.
Concentration:
20,000 units/ml
Unit Assay Substrate:
λ DNA
Storage Conditions:
25 mM Tris-HCl
25 mM KCl
1 mM Dithiothreitol
0.5 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.5 @ 25°C
Storage Temperature:
-20°C
Diluent Compatibility:
Diluent A
Notes
General notes:- Inhibited by salt concentrations > 10mM.
FAQs
- Is PmlI affected by methylation?
- Is PmlI inhibited by salt?
- Are storage conditions other than-20°C recommended?
- Is overnight digestion with PmlI recommended?
- Can PmlI be diluted?
- What is the activity of PmlI at 25°C?
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 PmlI, > 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 PmlI.
16-Hour Incubation:
A 50 μl reaction containing 1 μg of λDNA
and 100 units of PmlI 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 PmlI 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 100 units of PmlI with 1 μg of
ΦX174 RF I DNA for 4 hours at 37ºC resulted
in < 10% conversion to RFII as determined by agarose gel electrophoresis.
Reagents Sold Separately
NEBuffer 1
BSA
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