BLNI-RO
Roche
Bln I (Avr II)
from Brevibacterium linens
About This Item
biological source
bacterial (Brevibacterium linens)
Quality Level
form
solution
specific activity
10000 U/mL
packaging
pkg of 1,000 U (11558170001 [10 U/μl])
pkg of 200 U (11558161001 [10 U/μl])
manufacturer/tradename
Roche
parameter
37 °C optimum reaction temp.
color
colorless
pH
8.1 (39 °F)
solubility
water: miscible
suitability
suitable for molecular biology
application(s)
life science and biopharma
sample preparation
foreign activity
Endonucleases, none detected (up to 20 U with MWM II-DNA)
Endonucleases, none detected (up to 20U with pBR 322-DNA)
shipped in
dry ice
storage temp.
−20°C
General description
Compatible ends
Bln I ends are compatible with ends generated by Nhe I, Spe I and Xba I.
Isoschizomers
Bln I is an isoschizomer of Avr II.
Note: The complete 13 site Avr II restriction map of the E.coli genome has been reported.
Methylation sensitivity
The enzyme is not known to be affected by methylation.
Specificity
CCTAGG
Restriction site: C↓CTAGG
C↓CTAGG
Heat inactivation: No inactivation of Bln I after incubation at 65 °C for 15 minutes.
Quality
1 μg λDNA is incubated for 16 hours in 50 μl SuRE/Cut Buffer H with an excess of Bln I. The number of enzyme units which do not change the enzyme-specific pattern is stated in the certificate of analysis.
Absence of exonuclease activity
Approximately 5 μg [3H] labeled calf thymus DNA are incubated with 3 μl Bln I for 4 hours at +37°C in a total volume of 100 μl 50 mM Tris-HCl, 10 mM MgCl2, 1 mM Dithioerythritol, pH approximately 7.5. Under these conditions, no release of radioactivity is detectable, as stated in the certificate of analysis.
Typical ligation and recutting assay
Bln I fragments obtained by complete digestion of 1 μg λ × EcoR I DNA ligated for 16 hours at +4°C with 1 U T4 DNA Ligase in 10 μl buffer that contains 66 mM Tris-HCl, 5 mM MgCl2, 5 mM Dithiothreitol, 1 mM ATP, pH 7.5 (at +20°C). The percentages of product that can be ligated and subsequently recut with Bln I and EcoR I (yielding the typical pattern of λ × EcoR I × Bln I fragments) are stated under "Lig" and "Rec" in the certificate of analysis.
DNA Profile
- λ: 2
- φX174: 0
- Ad2: 2
- M13mp7: 0
- M13mp18:0
- pBR322: 0
- pBR328: 0
- pUC18: 0
- SV40: 2
Unit Definition
Storage and Stability
Analysis Note
Bln I has been tested in Pulsed-Field Gel Electrophoresis (on bacterial chromosomes). For cleavage of genomic DNA (E.coli C 600) embedded in agarose for PFGE analysis, we recommend using 10 U of enzyme/μg DNA and 4 hour incubation.
The buffer in bold is recommended for optimal activity
- A: 25-50%
- B: 50-75%
- H: 100%
- L: 0-10%
- M: 25-50%
Other Notes
Kit Components Only
- Enzyme Solution
- SuRE/Cut Buffer H 10x concentrated
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
does not flash
Flash Point(C)
does not flash
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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