Transfected Stable Cell Lines
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Cat. No. : EROT0315
| Cat. No. | EROT0315 |
| Description | Clostridium formicoaceticum. |
| Concentration | 10u/μl |
| Recognition Site | G CG↓C |
| Size | 3000 units |
| Reaction Conditions | Buffer B. 37 °C. |
| Storage | 10mM Tris-HCl (pH 7.4), 50mM NaCl, 0.1mM EDTA, 1mM DTT, 0.5mg/ml BSA, 50% glycerol. |
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.
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.
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.
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.
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.
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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