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FastDigest Bpu10I

5'...C C↓T N A G C...3'
3'...G G A N T↑C G...5'

FastDigest buffer for 100% activity Optimal incubation at 37°C Thermal inactivation at 80°C in 5 min Recombinant enzyme LO certified

The FastDigest Bpu10l restriction enzyme recognizes CCTNAGC(-5/-2)^ sites and cuts best at 37°C in 5-15 minutes using universal Yellow Tango Buffer
  
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Lambda DNA digested with Bpu10I

Lambda DNA digested with Bpu10I, 0.7% agarose, 4 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

For cleavage with FastDigest Bpu10I, at least two copies of its recognition sequence are required. DNA substrates with one site are cleaved about five times slower. Complete cleavage of some substrates by FastDigest Bpu10I is difficult to achieve.

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.

  
Storage Condition-20 C
HazardousNo
Compatible EndsFastDigest BlpI (Bpu1102I), FastDigest Bsu36I (Eco81I), FastDigest DdeI (HpyF3I), Bpu1102I, HpyF3I, Eco81I.
Formulation1 μL of enzyme (1 FDU) cleaves 1 μg of M13mp18 DNA in 15 minutes at 37°C in 1X FastDigest Buffer.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Recommended Reaction Conditions

1X FastDigest buffer or 1X FastDigest Green buffer at 37°C.
1 µL of FastDigest Bpu10I is formulated to digest up to:

  • 1 µg of M13mp18 DNA in 15 minutes
  • 1 µg of plasmid DNA in 15 minutes
  • 0.2 µg of PCR product in 30 minutes
  • 1 µg of genomic DNA in 15 minutes or 5 µg of genomic DNA in 60 minutes

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 uL Plasmid DNA, 1 µg/20 µL PCR product, ~0.2 µg/30 µL Genomic DNA, 1 µg/10 µL
37°C 15
(M13mp18 DNA)		 15 30 15 3 80°C, 5 min 1

Methylation Effects

Methylation type Sequence Cleavage effect
CpG5'...CCTNAGm5C G...3'
3'...GGANTC Gm5C...5'		 No effect
EcoBI(TGA(N)8TGCT)5'...CCTGm6AGC(N)6 TGCT...3'
3'...GGAC TCG(N)6m6ACGA...5'		 No effect

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
19 7 4 1 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 5 3

New sites generated by ligation

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

Recognition SequenceNewly generated sequence after reactionEnzymes that cleave the newly generated sequence
CC^TCAGCCCTCATCAGC
  • [MnlI/FastDigest MnlI]