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#S1 nuclease

2 public questions tagged with this topic.

Transcript analysis

Which of the following statements is true for S1 endonuclease?

S1 nuclease, purified from Aspergillus oryzae, is a single-strand specific endonuclease widely used in transcript analysis. It hydrolyzes phosphodiester bonds in single-stranded DNA and RNA, degrading unpaired regions. Double-stranded DNA with nicks, gaps, or loops also serves as substrate because local single-stranded character is exposed. It does not cleave fully base-paired duplexes. This property underpins S1 nuclease protection assay, where RNA-DNA hybrids are protected while unhybridized single-stranded tails are digested, allowing precise mapping of transcript termini and intron-exon bo

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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RDT Basics

S1 nuclease cleaves:

S1 nuclease is a zinc-dependent acidic endonuclease purified from the filamentous fungus Aspergillus oryzae. It exhibits high specificity for single-stranded nucleic acids, degrading unpaired DNA and RNA into 5'-mononucleotides while leaving fully base-paired duplexes largely intact under titrated conditions. Its activity requires zinc ions and low pH around 4.5. Molecular biologists utilize S1 nuclease to remove single-stranded overhangs after restriction digestion to create blunt ends, to cleave hairpin loops formed during second-strand cDNA synthesis, and to map transcription start sites an

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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