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FaqI (BsmFI) 

5'-G G G A C (N)10^-3'
3'-C C C T G (N)14^-5'

digested lambda DNA
1.0% agarose
       
#ER1811
 Supplied with:
10X Buffer Tango™
50X SAM		 100 u
 
1 ml
0.1 ml
Commercial Isoschizomers: BslFI, BsmFI

Related Documents (in pdf, ~55 KB):

Certificate of Analysis: #ER1811
MSDS (English)
MSDS (English-USA)
MSDS (German)
Chart (in pdf, 721 KB)
Brochure (in pdf, 316 KB)

Concentration
2 u/µl

Conditions for 100% Activity

Recommended
buffer		 Incubation
temperature		 Units for 
overnight 
incubation,
u/µg DNA		 Thermal
inactivation,
in 20 min

Enzyme activity, %
B 
(blue)		 G 
(green)		 O 
(orange)		 R 
(red)		 Tango™ 
(yellow)
1X 1X 1X 1X 1X 2X
Buffer Tango™ + 
0.05 mM S-adenosylmethionine		 37°C 1.0 80°C 20-50
(+SAM)		 20-50
(+SAM)		 0-20
(+SAM)		 0-20
(+SAM)		 100
(+SAM)		 20-50
(+SAM)

Storage Buffer
10 mM Tris-HCl (pH 7.5 at 25°C), 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/ml BSA and 50% glycerol.

Ligation and Recleavage
After 10-fold overdigestion with FaqI, more than 90% of the digested DNA fragments can be ligated but none of these can be recut due to the methylation of the recognition sequence by restriction enzyme.

Methylation Effects
Dam:
Dcm:
CpG:
EcoKI:
EcoBI:		  never overlaps
 may overlap
 may overlap
 never overlaps
 may overlap		 - no effect.
- no effect.
- blocked.
- no effect.
- no effect.

Note

  • FaqI requires only Mg2+ for its activity, but is stimulated by S-adenosylmethionine. 0.05 mM S-adenosylmethionine gives more than a 2-fold increase in FaqI activity. Still, a complete cleavage of some substrates is difficult to achieve.

  • FaqI concentration is determined by a maximal cleavage level achieved when no change in the fragmentation patterns is observed with the further enzyme increase.

  • FaqI may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid an atypical DNA band pattern, use the 6X Loading Dye & SDS Solution (#R1151) for sample preparation or heat the digested DNA in the presence of SDS prior to electrophoresis.

Double Digestions using Tango™ Buffer, DoubleDigest™

Optimal
buffer

Enzyme activity, %
Tango™ 
(yellow)
1X 2X
Buffer Tango™ + 
0.05 mM S-adenosylmethionine		 100
(+SAM)		 20-50
(+SAM)

Number of Recognition Sites in DNA Molecules

Lambda PhiX174 M13mp18/19 pBR322 pUC18/19 pUC57 pTZ19R/U pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
38 2 2 4 0 0 0/1 0/1 0/1 1 7

See also General Properties of Restriction Enzymes:

 

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Updated kovo 05, 2008 16:48