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Highly Modified Synthetic mRNA








Applications:
  • High efficiency protein synthesis in vitro by translation in rabbit reticulocyte lysate
  • High efficiency of protein synthesis in living cells, plus a bypass of innate antiviral responses
  • In vitro Diagnostics

Modifications include 5’ terminal modifications with cap, internal modification such as 5-methylcytidine or pseudouridine and 3’ modification such as poly A tail.


RNA
THE SERVICE PROVIDED BY AMSBIO


The Full Service includes:

Synthesis of synthetic DNA, plasmid cloning, purification, full DNA sequencing documentation, PCR amplification of insert, purification and characterisation with subsequent RNA synthesis by in vitro transcription, purification and characterisation. The customer provides text files with the desired sequence data.




Production Steps:
  1. Chemical synthesis of dsDNA fragments
  2. Cloning of the DNA fragments in a plasmid
  3. Quality control by sequencing of plasmid inserts
  4. Synthesis of PCR primers, based on the plasmid sequence, for PCR amplification of the cloned insert sequence. If desired, a 3’-Poly(A) tail can be included
  5. PCR amplification of the cloned insert with the primers described in step 4
  6. Purification of the PCR products with spin columns
  7. Quality control of PCR products by capillary electrophoresis (Agilent Bioanalyzer)
  8. PCR templates for production of ssRNA by in vitro transcription with T7 RNA polymerase
  9. DNase treatment for removal of the DNA template
  10. Purification of ssRNA with spin columns
  11. Quality control of the in vitro transcripts by capillary electrophoresis (Agilent Bioanalyzer)

mRNA can be made as research grade or GMP grade product.



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