CfoI

Name: CfoI
SKU: EROT0315

For research use only. Not intended for any clinical use.

Cat.No.

EROT0315

Description

Clostridium formicoaceticum.

Concentration

10u/μl

Reaction Conditions

Buffer B. 37 °C.

Recognition Site

G CG↓C

Size/Form

3000 units

Storage

10mM Tris-HCl (pH 7.4), 50mM NaCl, 0.1mM EDTA, 1mM DTT, 0.5mg/ml BSA, 50% glycerol.

Publications

Q & A

Customer Reviews

Customer Q&As

How does the recognition sequence specificity of CfoI compare to other commonly used restriction enzymes in molecular cloning procedures?

A: CfoI recognizes and cleaves the DNA sequence GCG|C. This 4-base palindromic recognition sequence is unique to CfoI and is distinct from other commonly used restriction enzymes, offering a specific cleavage option in molecular cloning procedures.

In the context of bisulfite sequencing, how can CfoI be used to determine methylation patterns at its specific recognition sites?

A: CfoI, like other restriction enzymes, is sensitive to DNA methylation at its recognition site. If a GCGC sequence is methylated, CfoI may not cleave it efficiently. Thus, by digesting bisulfite-treated DNA with CfoI and then sequencing, researchers can identify whether the specific GCGC sites were originally methylated.

Are there any known isoschizomers of CfoI, and if so, do they exhibit the exact same cleavage patterns and buffer preferences?

A: Isoschizomers are enzymes from different bacterial species that recognize the same DNA sequence. To date, there might be other enzymes with similar recognition sequences, but they could have varying cleavage patterns or buffer preferences. Researchers should refer to the most updated enzyme databases or catalogs for specific details.

How does the fidelity of CfoI cleavage get affected under non-optimal reaction conditions, such as suboptimal temperatures or ionic conditions?

A: Like many restriction enzymes, CfoI may exhibit star activity, or nonspecific cleavage, under non-optimal conditions. Factors like high enzyme concentration, non-ideal temperature, or incorrect buffer conditions can cause CfoI to cleave sites other than its primary GCG|C recognition sequence.

How efficient is CfoI in digesting DNA with secondary structures or when the recognition site is in close proximity to the end of a DNA fragment?

A: While CfoI is generally efficient at cleaving its recognition sequence, DNA secondary structures or sites too close to the fragment ends may impede its access or cleavage efficiency. It's advisable to provide ample flanking regions around the desired cut site to ensure complete digestion.

Can CfoI be used in conjunction with nicking enzymes to produce single-stranded overhangs, and if so, what potential applications does this have in DNA assembly or other molecular biology techniques?

A: Yes, CfoI can be paired with nicking enzymes that recognize adjacent or nearby sites to produce DNA fragments with single-stranded overhangs. This can facilitate techniques like Gibson Assembly or other methods where ssDNA overhangs can aid in the precise joining of DNA fragments.

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