FastDigest® MscI (MlsI)

Kód produktu: FD1214 Kód výrobce: FD1214 Kód dodavatele: {2D9F587D-EB43-4DBF-ACB9-B316EAF4E1E4} Výrobce: UAB Fermentas
2 795,10 Kč
2 310,00 Kč bez DPH
do týdne
50 rxns

5'...T G GC C A...3'
3'...A C CG G T...5'

Zobraz detailní popis

Detailní popis

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.

Formulation 1 μL of enzyme (1 FDU) cleaves 1 μg of lambda DNA dcm- in 5 minutes at 37°C in 1X FastDigest Buffer.
Hazardous No
Isoschizomers Search for commercial isoschizomers using REsearch.
Recommended Reaction Conditions

1X FastDigest buffer or 1X FastDigest Green buffer at 37°C.
1 µL of FastDigest MscI (MlsI) is formulated to digest up to:

  • 1 µg of lambda DNA dcm- in 5 minutes
  • 1 µg of plasmid DNA in 5 minutes
  • 0.2 µg of PCR product in 30 minutes
  • 1 µg of genomic DNA in 20 minutes or 5 µg of genomic DNA in 60 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 30 20 3 80°C, 20 min 16

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: may overlap - blocked.
  • CpG: never overlaps - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: never overlaps - no effect.
Methylation type Sequence Cleavage effect
Dcm(CCWGG) 5'...TGGCm5CA GG...3'
3'...ACCG GTm5CC...5'
Blocked

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
18 0 1 1 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 2

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
TGG^CCA
  • AluI/FastDigest AluI (AG^CT)
  • CviJI* (RG^CT)
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • Eco47III (AfeI)/FastDigest AfeI (Eco47III) (AGC^GCT)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)* (CCG^CTC)
  • MspA1I* (CMG^CTG)
  • PvuII/FastDigest PvuII (CAG^CTG)
  • CviJI
  • Bst1107I (BstZ17I)/FastDigest BstZ17I (Bst1107I) (GTA^TAC)
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI) (GG^CC)
  • CviJI* (RG^CC)
  • Eco147I (StuI)/FastDigest StuI (Eco147I) (AGG^CCT)
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • CviJI
  • Eco32I (EcoRV)/FastDigest EcoRV (Eco32I) (GAT^ATC)
  • Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)
  • BspPI (AlwI)
  • DpnI/FastDigest DpnI
  • MboI/FastDigest MboI
  • EheI (SfoI)/FastDigest EheI (GGC^GCC)
  • BmgT120I
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • Cfr13I (Sau96I)/FastDigest Sau96I (Cfr13I)
  • CviJI
  • FspAI/FastDigest FspAI* (RTGC^GCAC)
  • BseSI (Bme1580I)/FastDigest Bme1580I (BseSI)
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • HincII (HindII)/FastDigest HincII* (GTY^GAC)
  • FaqI (BsmFI)/FastDigest BsmFI (FaqI)

Obsah balení

50 rxns

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