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BLNI-RO

Roche

Bln I (Avr II)

from Brevibacterium linens

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Synonym(s):
restriction enzyme

biological source

bacterial (Brevibacterium linens)

Quality Level

form

solution

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.

storage temp.

−20°C

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This Item
XBAI-ROSMAI-RONOTI-RO
Bln I (Avr II) from Brevibacterium linens

Roche

BLNI-RO

Bln I (Avr II)

Xba I from Xanthomonas campestris

Roche

XBAI-RO

Xba I

Sma I from Serratia marcescens Sb

Roche

SMAI-RO

Sma I

Not I from Nocardia otitidis-caviarum

Roche

NOTI-RO

Not I

form

solution

form

solution

form

solution

form

solution

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Roche

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

Quality Level

100

Quality Level

100

Quality Level

100

Quality Level

100

packaging

pkg of 1,000 U (11558170001 [10 U/μl])

packaging

pkg of 1,000 U (10674257001 [10 U/μl]), pkg of 20,000 U (10674273001 [10 U/μl]), pkg of 20,000 U (11047663001 [40 U/μl]), pkg of 5,000 U (10674265001 [10 U/μl])

packaging

pkg of 1,000 U (10220566001 [10 U/μl]), pkg of 5,000 U (10656348001 [10 U/μl]), pkg of 5,000 U (11047639001 [40 U/μl])

packaging

pkg of 1,000 U (11014714001 [10 U/μl]), pkg of 1,000 U (11037668001 [40 U/μl]), pkg of 200 U (11014706001 [10 U/μl])

General description

Bln I recognizes the sequence C↓CTAGG 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: C↓CTAGG
C↓CTAGG
Heat inactivation: No inactivation of Bln I after incubation at 65 °C for 15 minutes.

Quality

Absence of nonspecific endonuclease activities
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

Number of cleavage sites on different DNAs
  • λ: 2
  • φX174: 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 λ 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

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.
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

Other Notes

For life science research only. Not for use in diagnostic procedures.

Kit Components Only

Product No.
Description

  • 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


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