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FastDigest BseSI

5'...G K G C MC...3'
3'...CM C G K G...5'

FastDigest buffer for 100% activity Optimal incubation at 37°C Thermal inactivation at 80°C in 15 min LO certified

The FastDigest BseSI restriction enzyme recognizes GKGCM^C sites and cuts best at 55°C in 5-15 minutes using universal FastDigest Buffer. This enzyme is an isoschizomer of Bme1580I having the same recognition and cleavage specificity (Isoschizomers: BaeGI, BstSLI).
  
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Lambda DNA digested with Bme1580I

Lambda DNA digested with BseSI, 0.7% agarose, 10 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.

  
Compatible Ends

 

 

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 BseSI 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 30 minutes or 5 µg of genomic DNA in 3 hours
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 30 3 80°C, 15 min 16

Methylation Effects

Methylation type Sequence Cleavage effect
Dcm(CCWGG)5'...GKGCCm5CW GG...3'
3'...CMCGG GWm5CC...5'
No effect
CpG5'...m5C GKGCMm5C G...3'
3'... Gm5CMCGK Gm5C...5'
No effect
EcoKI(AAC(N)6GTGC)5'...Am6AC(N)6G TGCMC...3'
3'...T TG(N)6Cm6ACGKG...5'
Impaired

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
10 1 2 3 3 4
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
2 3 3 3 2 1

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
GGGCA^C
  • SduI (Bsp1286I)/FastDigest SduI* (GGGCA^C)
  • BseSI (Bme1580I)/FastDigest BseSI
  • SduI (Bsp1286I)/FastDigest SduI
GGGCC^C
  • ApaI/FastDigest ApaI (GGGCC^C)
  • Eco24I (BanII)* (GGGCC^C)
  • SduI (Bsp1286I)/FastDigest SduI* (GGGCC^C)
  • ApaI/FastDigest ApaI
  • BmgT120I
  • BseSI (Bme1580I)/FastDigest BseSI
  • Bsp120I (PspOMI)/FastDigest Bsp120I
  • BspLI (NlaIV)/FastDigest BspLI
  • BsuRI (HaeIII)/FastDigest BsuRI
  • Cfr13I (Sau96I)/FastDigest Cfr13I
  • CviJI
  • Eco24I (BanII)
  • SduI (Bsp1286I)/FastDigest SduI
GTGCA^C
  • Alw21I (BsiHKAI)/FastDigest Alw21I* (GTGCA^C)
  • SduI (Bsp1286I)/FastDigest SduI* (GTGCA^C)
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • Alw44I (ApaLI)/FastDigest Alw44I
  • BseSI (Bme1580I)/FastDigest BseSI
  • Hpy8I (MjaIV)/FastDigest Hpy8I
  • HpyCH4V
  • SduI (Bsp1286I)/FastDigest SduI
  • Mph1103I (NsiI)/FastDigest Mph1103I (ATGCA^T)
  • HpyCH4V
  • PstI/FastDigest PstI (CTGCA^G)
  • SdaI (SbfI)/FastDigest SdaI (CCTGCA^GG)
  • BsgI
  • HpyCH4V
GTGCC^C
  • SduI (Bsp1286I)/FastDigest SduI* (GTGCC^C)
  • BseSI (Bme1580I)/FastDigest BseSI
  • SduI (Bsp1286I)/FastDigest SduI

 

References

Citations