Viswagen Biotech Pvt. Ltd.

Alphabetic List of SBS Restriction Endonucleases

Restriction Endonucleases (N-R)

Nae I

GCC

GGC

Cat. #	Size	Conc.
122-1	150 units	8-12u/µl

Source:	Streptomyces species

Buffer supplied:	10×L

Substrate for unit definition:	pBR322

Reaction conditions:	10 mM Tris-HCl (pH 7.9, 25^oC), 10 mM MgCl2, 1 mM dithiothreitol, 100 µg/ml BSA. Incubate at 37^oC.

Storage buffer:	50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Fifty units of Nae I do not produce any unspecific cleavage products after 16 hrs incubation with 1µg of Lambda DNA at 37^oC. After ten-fold overdigestion with Nae I, greater than 80% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65^oC for 20 minutes.

Note:	Nae I exhibits site preferences.

Nco I

CATGG

Cat. #	Size	Conc.
123-1	400 units	8-12u/µl

Source:	Nocardia corallina

Buffer supplied:	10×Nco I

Substrate for unit definition:	Lambda DNA

Reaction conditions:	100 mM NaCl, 50 mM Tris-HCl (pH 7.9, 25^oC), 10 mM MgCl2, 1 mM dithiothreitol, 0.02% Triton X-100, 100 µg/ml BSA. Incubate at 37^oC.

Storage buffer:	50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Fifty units of Nco I do not produce any unspecific cleavage products after 16 hrs incubation with 1µg of Lambda DNA at 37^oC. After ten-fold overdigestion with Nco I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65^oC for 20 minutes.

Star activity:	Conditions of high enzyme concentration or glycerol concentration>5%, may result in star activity.

Nhe I

CTAGC

Cat. #	Size	Conc.
146-1	150 units	8-12u/µl

Source:	Neisseria mucosa heildelbergensis (ATCC 25999)

Buffer supplied:	10×A

Substrate for unit definition:	Lambda DNA (Hind III digest)

Reaction conditions:	50 mM potassium acetate, 20 mM Tris-acetate (pH 7.9, 25^oC), 10 mM magnesium acetate, 1 mM dithiothreitol, 100 µg/ml BSA. Incubate at 37^oC.

Storage buffer:	50 mM KCl, 10 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM dithiothreitol, 200µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Forty units of Nhe I do not produce any unspecific cleavage products after 16 hrs incubation with 1µg of Lambda DNA at 37^oC. After 50-fold overdigestion with Nhe I, greater than 98% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65^oC for 20 minutes.

Star activity:	Large excess of the enzyme may results in the appearance of star activity.

Note:	Activity inhibited by salt concentractions >100mM. Cleaves to leave a 5'CTAG extension which can be efficiently ligated to DNA fragments generated by Avr II, Spe I, or Xba I.

Not I

GGCCGC

Cat. #	Size	Conc.
124-1	200 units	8-12u/µl

Source:	Nocardia otitidis-caviarum

Buffer supplied:	10×Not I

Substrate for unit definition:	Adenovirus-2 DNA

Reaction conditions:	100 mM NaCl, 50 mM Tris-HCl (pH 7.9, 25^oC), 5 mM MgCl2, 1mM dithiothreitol. Incubate at 37^oC.

Storage buffer:	500 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1mM dithiothreitol, 0.1% Triton X-100, 500 µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Fifty units of Not I do not produce any unspecific cleavage products after 16 hrs incubation with 1µg of Adeno-2 DNA at 37^oC. After 30-fold overdigestion with Not I, greater than 98% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65^oC for 20 minutes.

Note:	Supercoiled plasmids may require up to 5-fold more Not I for complete digestion than linear DNAs

Nru I

TCG

CGA

Cat. #	Size	Conc.
125-1	2,00 units	8-12u/µl

Source:	Nocardia rubra

Buffer supplied:	10×Nru I

Substrate for unit definition:	Lambda DNA

Reaction conditions:	100 mM KCI, 50 mM Tris-HCl (pH 8.0, 25^oC), 10 mM MgCl2, 100µg/ml BSA. Incubate at 37^oC.

Storage buffer:	50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Fifty units of Nru I do no produce any unspecific cleavage products after 16 hrs incubation with 1 µg of Lambda DNA at 37^oC. After ten-fold overdigestion with Nru I, less than 20% of the DNA fragments can be ligated.

Heat inactivation:	65 ^oC for 20 minutes.

Star activity:	Large excess of the enzyme results in the appearance of star activity.

PspP I (Sau96 I, Cfr13 I)

GNCC

Cat. #	Size	Conc.
126-1	1,000 units	8-12u/µl

Source:	Psychrobacter immobilis TA137

Buffer supplied:	10×M

Substrate for unit definition:	Lambda DNA

Reaction conditions:	50 mM NaCl, 10 mM Tris-HCl (pH 7.9, 25^oC), 10 mM MgCl2, 1 mM DTT,100 µg/ml BSA, Incubate at 25^oC.

Storage buffer:	50 mM NaCl,10mM Tris-HCl (pH 7.4, 25^oC),0.1 mM EDTA, 1 mM DTT,200 µg/mlBSA, 50%glycerol. Store at -20^oC.

Absence of contaminants:	Fifty units of PspP I do not produce any unspecific cleavage products after 16 hrs incubation with 1 µg of Lambda DNA at 37^oC. After ten-fold overdigestion with PspP I, greater than 95% of the DNA fragments can be ligated.

Heat inactivation:	55^oC for 15minutes.

Pst I

CTGCA

Cat. #	Size	Conc.
127-1	5000 units	8-12u/µl

Source:	An E. coli strain that carries the cloned Pst I gene from Providencia stuartii

Buffer supplied:	10xPst I

Substrate for unit definition:	Lambda DNA

Reaction conditions:	100 mM NaCl, 50 mM Tris-HCl (pH 7.4, 25^oC), 10 mM MgCl2, 1 mM dithiothreitol, 100 µg/ml BSA. Incubate at 37^oC.

Storage buffer:	200 mM NaCl, 10mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 µg/ml BSA, 0.15% Triton X-100 and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Ten units of Pst I do not produce any unspecific cleavage products after 16 hrs incubation with 1 µg of Lambda DNA at 37^oC. After ten-fold overdigestion with Pst I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	80 ^oC for 20 minutes.
Star activity:	Conditions of high enzyme concentration or glycerol concentration>12% may result in star activity.

Pvu II

CAG

CTG

Cat. #	Size	Conc.
128-1	2,000 units	8-12u/µl

Source:	An E. coli strain that carries the cloned Pvu II gene from Proteus vulgaris

Buffer supplied:	10xM

Substrate for unit definition:	Lambda DNA

Reaction conditions:	50 mM NaCl, 10 mM Tris-HCl (pH 7.9, 25^oC), 10 mM MgCl2, 1 mM dithiothreitol, 100 µg/ml BSA. Incubate at 37^oC.

Storage buffer:	50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Ten units of Pvu II do not produce any unspecific cleavage products after 16 hrs incubation with 1 µg of Lambda DNA at 37^oC. After ten-fold overdigestion with Pvu II, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	No.

Star activity:	Conditions of low ionic strength, high enzyme concentration, glycerol concentration>5%, or pH>8.0 may result in star activity.

Rsa I(Afa I)

Cat. #	Size	Conc.
129-1	500 units	8-12u/µl

Source:	Rhodopseudomonas sphaeroides

Buffer supplied:	10×M

Substrate for unit definition:	Lambda DNA

Reaction conditions:	50 mM NaCl, 10 Tris-HCl (pH 7.9, 25^oC), 10 mM MgCl2, 1 mM dithiothreitol, 100 µg/ml BSA. Incubate at 37^oC.

Storage buffer:	50 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Forty units of Rsa I do not produce any unspecific cleavage products after 16 hrs incubation with 1 µg of Lambda DNA at 37^oC. After ten-fold overdigestion with Rsa I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65^oC for 20 minutes.

Note:	Cleaves single-strand DNA slowly

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