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SspDI (KasI)

Kód produktu: ER2191 Kód výrobce: ER2191 Kód dodavatele: {BEC1DBC3-33B0-4059-B933-D02E22428E3B} Výrobce: Life Technologies Czech Republic s.r.o.
2 674,10 Kč
2 210,00 Kč bez DPH
do týdne
250 Units

5'...GG C G C C...3'
3'...C C G C GG...5'

Zobraz detailní popis

Detailní popis

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Highlights

  • Superior quality – stringent quality control and industry leading manufacturing process
  • Convenient color-coded Five Buffer System
  • Includes universal Tango buffer for double-digestions
  • BSA premixed in reaction buffers
  • Wide selection of restriction endonuclease specificities

Applications

  • Molecular cloning
  • Restriction site mapping
  • Genotyping
  • Southern Blot
  • Restriction fragment length polymorphism (RFLP)
  • SNP

Note

For methylation sensitivity refer to product specifications.

Compatible Ends BshNI (BanI).
Conditions for 100% Activity
  • 1X Buffer Tango: 33 mM Tris-acetate (pH 7.9 at 37°C), 10 mM Mg-acetate, 66 mM K-acetate, and 0.1 mg/mL BSA
  • Incubate at 37°C
Double Digestion Perform double digestion using DoubleDigest.
Hazardous No
Isoschizomers Search for commercial isoschizomers using REsearch.
Note Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).
Storage Buffer SspDI is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM KCl, 1 mM DTT, 1 mM EDTA, 0.2 mg/mL BSA, and 50% (v/v) glycerol
Storage Condition -20 C

Reaction conditions

Recommended buffer for 100% activity Optimal temperature Enzyme activity in Thermo Scientific buffers, % Tango buffer for double digestion
B (blue)
1X
G (green)
1X
O (orange)
1X
R (red)
1X
Tango
(yellow)
1X / 2X
Tango 37°C; 20-50 0-20 NR 0-20 100 20-50 1X or 2X

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: may overlap - no effect.
  • CpG: completely overlaps - blocked.
  • EcoKI: never overlaps - no effect.
  • EcoBI: never overlaps - no effect.
Methylation type Sequence Cleavage effect
Dcm (CCWGG) 5'...Cm5CW GGCGCm5CW GG...3'
3'...G GWm5CCGCG GWm5CC...5'
No effect
CpG GGCGCC Blocked

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
20-50 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
1 2 1 4 1 1
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 4

New sites generated by ligation

Newly generated recognition sites resulting from ligation of protruding compatible DNA ends

Recognition Sequence Second Enzyme Enzymes recognizing newly generated recognition sequence
G^GCGCC
  • BshNI (BanI)/FastDigest BanI (BshNI)* (G^GCGCC)
  • BbeI
  • FastDigest HaeII (BfoI)
  • BshNI (BanI)/FastDigest BanI (BshNI)
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • EheI (SfoI)/FastDigest EheI
  • HhaI/FastDigest HhaI
  • Hin1I (BsaHI)/FastDigest BsaHI (Hin1I)
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • NarI
  • SspDI (KasI)

Newly generated recognition sites resulting from fill-in of 5'-overhang and self-ligation

Recognition Sequence Newly generated sequence after reaction Enzymes that cleave the newly generated sequence
G^GCGCC GGCGCGCGCC
  • Bsh1236I (BstUI)/FastDigest Bsh1236I
  • Cac8I
  • [HhaI/FastDigest HhaI]
  • [Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)]
  • PauI (BssHII)/FastDigest BssHII (PteI)

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