Viswagen Biotech Pvt. Ltd.

Alphabetic List of SBS Restriction Endonucleases

Restriction Endonucleases (S-X)

Sal I

TCGAC

Cat. #	Size	Conc.
130-1	2,500 units	8-12u/µl

Source:	Streptomyces albus G

Buffer supplied:	10×SH

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

Reaction conditions:	150 mM NaCl, 10 mM Tris-HCl (pH 7.9, 25^oC), 10 mM MgCl2, 1 mM dithiothreitol, 100µg/ml BSA. Incubate at 37^oC.

Storage buffer:	50 mM KCl, 5 mM KPO4 (pH 7.5), 0.1 mM EDTA, 1 mM dithiothreitol, 200µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Ten units of Sal I do not produce any unspecific cleavage products after 16 hrs incubation with 1µg of Lambda DNA (Hind III digest) at 37^oC. After ten-fold overdigestion with Sal 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	Large excess of the enzyme results in the appearance of star activity.

Sau3A I (Mob I)

GATC

Cat. #	Size	Conc.
147-1	200 units	8-12u/µl

Source:	Streptomyces species

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. Incubate at 37^oC.

Storage buffer:	50 mM KCl, 10 mM Tris-HCl (pH 7.4, 25^oC), 0.1 mM EDTA, 1 mM dithiothreitol, 200µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Forty units of Sau3A 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 Sau3A I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65^oC for 20 minutes.

Note:	Sau3A I and Mob I are isoschizomers but Sau3A I, unlike Mbo I, is not blocked by dam methylation. That means that Sau3A I cuts the sequence GATC when A is methylated (GmATC) but Mbo I dose not.

Sca I

AGT

ACT

Cat. #	Size	Conc.
131-1	500 units	8-12u/µl

Source:	Streptomyces caespitosus

Buffer supplied:	10×Sca I

Substrate for unit definition:	Lambda DNA

Reaction conditions:	100 mM NaCl, 10 mM Tris-HCl (pH 7.4, 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:	Fifty units of Sca 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 Sca I, greater than 90% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	80^oC for 20 minutes.

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

Sfi I

GGCCNNNN

NGGCC

Cat. #	Size	Conc.
132-1	500 units	8-12u/µl

Source:	Streptomyces fimbriatus

Buffer supplied:	10×M

Substrate for unit definition:	Adenovirus-2 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 50^oC under parafin oil.

Storage buffer:	50 mM NaCl, 10 mM Tris-HCl (pH 7.9, 25^oC), 10 mM MgCl2, 1 mM dithiothreitol, 100 µg/ml BSA. Incubate at 50^oC under parafin oil.

Absence of contaminants:	Fifty units of Sfi I do not produce any unspecific cleavage products after 16 hrs incubation with 1µg of Adeno-2 DNA at 50^oC. After ten-fold overdigestion with Sfi I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	No

SgrB I (Sac II)

CCGC

Cat. #	Size	Conc.
133-1	1,000 units	8-12u/µl

Source:	Streptomyces griseus

Buffer supplied:	10×SgrB I

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

Reaction conditions:	10 mM Tris-HCl (pH 7.9, 25^oC), 10 mM MgCl2, 1 mM dithiothreitol, 0.1% Triton X-100, 100 µg/ml BSA. Incubate at 37^oC.

Storage buffer:	50 mM NaCl, 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 SgrB I do not produce any unspecific cleavage products after 16 hrs incubation with 1 µg of Lambda DNA (Hind III digest) at 37^oC. After ten-fold overdigestion with SgrB I, greater than 98% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65 ^oC for 20 minutes.

Note:	Particular sites in Lambda and Fi X174 DNAs are difficult to cleave with SgrB I, as well as with its prototype Sac II.

Sla I (Xho I)

TCGAG

Cat. #	Size	Conc.
134-1	2,500 units	8-12u/µl

Source:	Streptomyces lavendulae

Buffer supplied:	10×SH

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

Reaction conditions:	150 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, 200 µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Fifty units of Sla I do not produce any unspecific cleavage products after 16 hrs incubation with 1 µg of Lambda DNA (Hind III digest) at 37^oC. After ten-fold overdigestion with Sla I, greater than 98% of theLambda DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65^oC for 20 minutes.

Sma I

CCC

GGG

Cat. #	Size	Conc.
135-1	1,000 units	8-12u/µl

Source:	Serratia marcescens

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 25^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 Sma I do not produce any unspecific cleavage products after 16 hrs incubation with 1 µg of Lambda DNA at 25^oC. After ten-fold overdigestion with Sma I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65^oC for 20 minutes.

SnaB I

TAC

GTA

Cat. #	Size	Conc.
136-1	150 units	8-12u/µl

Source:	Sphaerotilus natans

Buffer supplied:	10×SnaB I

Substrate for unit definition:	Lambda DNA (EcoR I digest)

Reaction conditions:	10 mM Bis Tris Propane-HCI (pH 7.0, 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:	Ten units of SnaB I do not produce any unspecific cleavage products after 16 hrs incubation with 1 µg of Lambda DNA/EcoR I digest at 37^oC.After ten-fold overdigestion with SnaB I, greater than 98% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	80^oC for 20 minutes.

Star activity:	Star activity is observed at a greater than 160-fold overdigestion with SnaB I.

Sph I

GCATG

Cat. #	Size	Conc.
137-1	250 units	8-12u/µl

Source:	Streptomyces phaeochromogenes

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 dithiothreitol, 100 µg/ml BSA. Incubate at 37^oC.

Storage buffer:	100 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 400 µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Fifty units of Sph 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 Sph I, greater than 98% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65^oC for 20 minutes.

SseB I (Stu I)

AGG

CCT

Cat. #	Size	Conc.
138-1	500 units	8-12u/µl

Source:	Streptomyces species

Buffer supplied:	10×H

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

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 7.9, 25^oC), 0.1 mM EDTA, 1 mM dithiothreitol, 200 µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Fifty units of SseB I do not produce any unspecific cleavage products after 16 hrs incubation with 1 µg of Lambda DNA/Hind III digest at 37^oC. After 10-fold overdigestion with SseB I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65^oC for 20 minutes.

Ssp I

AAT

ATT

Cat. #	Size	Conc.
139-1	150 units	8-12u/µl

Source:	Sphaerotilus species

Buffer supplied:	10xH

Substrate for unit definition:	Lambda DNA

Reaction conditions:	100 mM NaCl, 50 mM Tris-HCl (pH 7.9, 25^oC), 10 mM MgCl2, 1mM dithiothreitol, 0.025% Triton X-100, 100µg/ml BSA. Incubate at 37^oC.

Storage buffer:	50 mM KCl, 10 mM Tris-HCl (pH 8.0), 0.1 mM EDTA, 1 mM dithiothreitol, 200µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Ten units of Ssp 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 Ssp 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 low ionic strength, high enzyme concentration, glycerol concentration >5%, or pH>8.0 may result in star activity.

Sst I (Sac I)

GAGCT

Cat. #	Size	Conc.
140-1	1,000 units	8-12u/µl

Source:	Streptomyces Stanford

Buffer supplied:	10×L

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

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 NaCl, 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 Sst I do not produce any unspecific cleavage products after 16 hrs incubation with 1µg of Lambda DNA/Hind III digest at 37^oC. After ten-fold overdigestion with Sst I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	65^oC for 20 minutes.

Sty I (EcoT14 I)

C(A/T)(T/A)GG

Cat. #	Size	Conc.
141-1	2,500 units	8-12u/µl

Source:	E.coli WA921/pST27 hsd+

Buffer supplied:	10×M

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

Heat inactivation:	65^oC for 20 minutes.

Taq I

CGA

Cat. #	Size	Conc.
142-1	1,000 units	8-12u/µl

Source:	Thermus aquaticus YT I

Buffer supplied:	10×Taq I

Substrate for unit definition:	Lambda DNA (dam-)

Reaction conditions:	100 mM KCl, 20 mM Tris-HCl (pH 8.5, 25^oC), 3 mM MgCl2, 0.04% Triton X-100. Incubate at 65^oC under parafin oil.

Storage buffer:	300 mM KCl, 10 mM Tris-HCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 500 µg/ml BSA and 50% glycerol. Store at -20^oC.

Absence of contaminants:	Fifty units of Taq I do not produce any unspecific cleavage products after 16 hrs incubation with 1µg of Lambda DNA at 65^oC. After 50-fold overdigestion with Taq I, greater than 90% of the DNA fragments can be ligated and recut with this enzyme.

Heat inactivation:	80^oC for 20 minutes.

Note:	Taq I is blocked by overlapping dam methylation. Incubation without BSA results in 50% activity.

Xba I

CTAGA

Cat. #	Size	Conc.
143-1	1,500 units	8-12u/µl

Source:	Xanthomonas badrii

Buffer supplied:	10×M

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

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

Storage buffer:	50 mM NaCl, 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:	Two hundred units of Xba I do not produce any unspecific cleavage products after 16 hrs incubation with 1µg of Lambda DNA at 37^oC. After 100-fold overdigestion with Xba 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:	Conditions of low ionic strength, high enzyme concentration, or glycerol concentration>5%, may result in star activity.

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