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FastDigest ScaI

5'...A G TA C T...3'
3'...T C AT G A...5'

FastDigest buffer for 100% activity Optimal incubation at 37°C Thermal inactivation at 65°C in 10 min LO certified

The FastDigest ScaI restriction enzyme recognizes AGT^ACT sites and cuts best at 37°C in 5-15 minutes using universal FastDigest Buffer (Isoschizomers: AssI, BmcAI, ZrmI)
  
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Lambda DNA digested with ScaI

Lambda DNA digested with ScaI, 0.7% agarose, 5 cleavage sites

Thermo Scientific FastDigest enzymes are an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.

The universal buffer allows rapid single-, double- or multiple DNA digestion within 5-15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.

For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.

Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.

Features

  • 100% activity of all FastDigest enzymes in the universal buffer
  • 100% buffer compatibility with downstream applications
  • Complete digestion in 5-15 minutes
  • Direct loading on gels
  • No star activity
  • 176 FastDigest enzymes available

Applications

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

Note

The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer. For applications that require product analysis by fluorescence excitation (e.g. concentration measurements in UV light) the colorless FastDigest Buffer is recommended.

For methylation sensitivity refer to product specifications.

  
Formulation1 µL of enzyme (1 FDU) cleaves 1 μg of lambda DNA in 5 minutes at 37°C in 1X FastDigest Buffer.
HazardousNo
IsoschizomersSearch for commercial isoschizomers using REsearch.
Recommended Reaction Conditions

1X FastDigest buffer or 1X FastDigest Green buffer at 37°C.
1 µL of FastDigest Sca is formulated to digest up to:

  • 1 µg of lambda DNA in 5 minutes
  • 1 µg of plasmid DNA in 5 minutes
  • 0.2 µg of PCR product in 5 minutes
  • 1 µg of genomic DNA in 5 minutes or 5 µg of genomic DNA in 30 minutes
Storage Condition-20 C

Reaction conditions

Reaction temperature Digestion time with 1 µL of FastDigest enzyme, min bp from end of DNA required for complete digestion Thermal inactivation Incubation time without star activity, hours
Lambda, 1 µg/20 µL Plasmid DNA, 1 µg/20 µL PCR product, ~0.2 µg/30 µL Genomic DNA, 1 µg/10 µL
37°C 5 5 5 5 4 65°C, 10 min 16

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: never overlaps - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: may overlap - blocked.
Methylation type Sequence Cleavage effect
EcoBI(TGA(N)8TGCT)5'...TGm6AGTACT(N)3 TGCT...3'
3'...AC TCATGA(N)3m6ACGA...5'
Blocked

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
5 0 0 1 1 1
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
1 1 1 1 1 1

New sites generated by ligation

Newly generated recognition sites resulting from ligation of blunt DNA ends

Recognition sequence Second restriction enzyme Restriction enzymes cleaving the newly generated recognition sequence
AGT^ACT
  • AanI (PsiI)/FastDigest PsiI (AanI) (TTA^TAA)
  •  
  • FastDigest MseI (SaqAI)
  • Tru1I (MseI)/FastDigest Tru1I
  • Bsp68I (NruI)/FastDigest NruI (RruI) (TCG^CGA)
  • TaqI/FastDigest TaqI
  • Bst1107I (BstZ17I)/FastDigest BstZ17I (Bst1107I) (GTA^TAC)
  • HincII (HindII)/FastDigest HincII* (GTY^AAC)
  • KspAI (HpaI)/FastDigest HpaI (KspAI) (GTT^AAC)
  • MaeIII
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • Alw26I (BsmAI)/FastDigest Alw26I
  • FspAI/FastDigest FspAI* (RTGC^GCAC)
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • Alw44I (ApaLI)/FastDigest ApaLI (Alw44I)
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)
  • Hpy8I (MjaIV)/FastDigest Hpy8I
  • HpyCH4V
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • FspAI/FastDigest FspAI* (RTGC^GCAT)
  • NsbI (FspI)/FastDigest FspI (NsbI) (TGC^GCA)
  • HpyCH4V
  • HincII (HindII)/FastDigest HincII* (GTY^GAC)
  • MaeIII
  • NmuCI (Tsp45I)/FastDigest NmuCI
  • MssI (PmeI)/FastDigest MssI (GTTT^AAAC)
  • Hpy8I (MjaIV)/FastDigest Hpy8I
  • RsaI/FastDigest RsaI (GT^AC)
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI

New sites generated by ligation

Newly generated recognition sites resulting from ligation of blunt DNA ends

Recognition sequence Second restriction enzyme Restriction enzymes cleaving the newly generated recognition sequence
ATGC^GCAY
  • DraI/FastDigest DraI (TTT^AAA)
  • HincII (HindII)/FastDigest HincII* (GTY^AAC)
  • KspAI (HpaI)/FastDigest HpaI (KspAI) (GTT^AAC)
  • MssI (PmeI)/FastDigest MssI (GTTT^AAAC)
  • RsaI/FastDigest RsaI (GT^AC)
  • ScaI/FastDigest ScaI (AGT^ACT)
  • SmiI (SwaI)/FastDigest SwaI (SmiI) (ATTT^AAAT)
  • HpyCH4VI
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • MnlI/FastDigest MnlI
  • Eco32I (EcoRV)/FastDigest EcoRV (Eco32I) (GAT^ATC)
  • LweI (SfaNI)/FastDigest SfaNI (BmsI)
  • HpyCH4V
  • Mph1103I (NsiI)/FastDigest NsiI (Mph1103I)
  • Eco47III (AfeI)/FastDigest AfeI (Eco47III) (AGC^GCT)
  • EheI (SfoI)/FastDigest EheI (GGC^GCC)
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (GAG^CGG)
  • MspA1I* (CMG^CGG)
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • NsbI (FspI)/FastDigest FspI (NsbI) (TGC^GCA)
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • NsbI (FspI)/FastDigest FspI (NsbI)
  • PdiI (NaeI)/FastDigest NaeI (PdiI) (GCC^GGC)
  • SatI (Fnu4HI)/FastDigest Fnu4HI (SatI)
  • TauI/FastDigest TauI
  • SrfI (GCCC^GGGC)
  • Cac8I
  • SspI/FastDigest SspI (AAT^ATT)
  • HpyCH4V
  • Mph1103I (NsiI)/FastDigest NsiI (Mph1103I)
GTGC^GCAY
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI) (GG^CC)
  • CviJI* (RG^CC)
  • Eco147I (StuI)/FastDigest StuI (Eco147I) (AGG^CCT)
  • MlsI (MscI)/FastDigest MscI (MlsI) (TGG^CCA)
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • DpnI/FastDigest DpnI (GA^TC)
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • DraI/FastDigest DraI (TTT^AAA)
  • HincII (HindII)/FastDigest HincII* (GTY^AAC)
  • KspAI (HpaI)/FastDigest HpaI (KspAI) (GTT^AAC)
  • MssI (PmeI)/FastDigest MssI (GTTT^AAAC)
  • SmiI (SwaI)/FastDigest SwaI (SmiI) (ATTT^AAAT)
  • SspI/FastDigest SspI (AAT^ATT)
  • HpyCH4V
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • MnlI/FastDigest MnlI
  • Eco32I (EcoRV)/FastDigest EcoRV (Eco32I) (GAT^ATC)
  • LweI (SfaNI)/FastDigest SfaNI (BmsI)
  • HpyCH4V
  • Eco47III (AfeI)/FastDigest AfeI (Eco47III) (AGC^GCT)
  • EheI (SfoI)/FastDigest EheI (GGC^GCC)
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (GAG^CGG)
  • MspA1I* (CMG^CGG)
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • NsbI (FspI)/FastDigest FspI (NsbI) (TGC^GCA)
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • NsbI (FspI)/FastDigest FspI (NsbI)
  • PdiI (NaeI)/FastDigest NaeI (PdiI) (GCC^GGC)
  • SatI (Fnu4HI)/FastDigest Fnu4HI (SatI)
  • TauI/FastDigest TauI
  • RsaI/FastDigest RsaI (GT^AC)
  • ScaI/FastDigest ScaI (AGT^ACT)
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • Alw44I (ApaLI)/FastDigest ApaLI (AIw44I)
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)
  • Hpy8I (MjaIV)/FastDigest Hpy8I
  • HpyCH4V
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • SrfI (GCCC^GGGC)
  • Cac8I

References

Citations