RNA Synthesis In vitro Transcription (IVT)
In vitro RNA synthesis refers to the synthesis of RNA from a DNA template using a phage DNA-dependent RNA polymerase (e.g. T7, T3 or SP6 RNA polymerase). Template DNA for
in vitro transcription (IVT) includes the RNA polymerase promoter upstream of the target sequence, and template options include plasmids, PCR products, oligonucleotides, and linearised DNA templates such as cDNA that contain the corresponding promoter.
In vitro transcripts are used in analytical techniques (e.g. hybridization analysis), structural studies (for NMR and X-ray crystallography), in biochemical and genetic studies (e.g. as antisense reagents), and as functional molecules (ribozymes and aptamers).
IVT products include RNA synthesis systems, nuclear extracts, ribonucleotides, cloning vectors and supporting products for transcription, primer extension and gene expression studies.
T7 RNA Polymerase is a DNA-dependent RNA polymerase with strict specificity for T7 promoter. Use double-stranded DNA (linearised plasmid or PCR product) as a template to synthesise RNA that is complementary to the reverse single-stranded DNA downstream of the promoter. T7 RNA Polymerase accepts modified nucleotides (e.g., biotin-, digoxigenin-, fluorescein-labeled nucleotides) as substrates for RNA synthesis.
DNase I is an endonuclease that digests single- and double- stranded DNA. It hydrolyzes phosphodiester bonds producing mono- and oligodeoxyribonucleotides with 5’-phosphate and 3’-OH groups. The enzyme activity is strictly dependent on Ca2+ and is activated by Mg2+ or Mn2+ ions. If in the presence of Mg2+, DNase I cleaves each strand of dsDNA independently, in a statistically random fashion. If in the presence of Mn2+, the enzyme cleaves both DNA strands at approximately the same site, producing DNA fragments with blunt-ends or with one or two nucleotide overhangs.
T7 RNA Synthesis Kit is designed for in vitro transcription of large amounts of RNA, provides with T7 RNA polymerase mix and four individually available nucleotides. The kit can also be used to synthesise capped RNA by adding cap analogues; or to synthesise modified RNA by replacing with modified UTP according to experimental requirements. The kit contains LiCl Solution for removing free nucleotides, enzymes and most of the template from the transcription product to obtain coarsely purified RNA.
T7 RNA Co-Transcription Synthesis Kit
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In-vitro Transcription Workflow
*To order the products above, please:
1) Place your order directly on our website.
2) Email your purchase order to sales@genecopoeia.com.
