Bln I (Avr II)

Кат. номер

11558170001

11558161001

Bln I recognizes the sequence CCTAGG and generates fragments with 5-cohesive termini.
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
Recognition sites: CCTAGG
CCTAGG
Restriction site: CCTAGG
CCTAGG
Heat inactivation: No inactivation of Bln I after incubation at 65 °C for 15 minutes.
Quality
Absence of nonspecific endonuclease activities
1 µg DNA 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 x 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 x EcoR I x Bln I fragments) are stated under "Lig" and "Rec" in the certificate of analysis.
DNA Profile
Number of cleavage sites on different DNAs

• : 2
• X174: 0
• Ad2: 2
• M13mp7: 0
• M13mp18:0
• pBR322: 0
• pBR328: 0
• pUC18: 0
• SV40: 2

Unit Definition
One unit is the enzyme activity that completely cleaves 1 µg x EcoR I DNA fragments in one hour at +37 °C in a total volume of 25 µl (1x) SuRE/Cut Buffer H.
Analysis Note
SuRE/Cut Buffer System
The buffer in bold is recommended for optimal activity

• A: 25-50%
• B: 50-75%
• H: 100%
• L: 0-10%
• M: 25-50%

Activity in PCR buffer: Not tested
PFGE tested
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.
For life science research only. Not for use in diagnostic procedures.

Quality Level	100,
form	solution
packaging	pkg of 1,000 U (11558170001 [10 U/µl]), pkg of 200 U (11558161001 [10 U/µl])
Торговая марка	Roche
parameter	37 °C optimum reaction temp.
shipped in	dry ice
storage temp.	20°C

RIDADR

NONH for all modes of transport