Method for improving the stability of linear DNA in cell-free in vitro transcription / translation systems
AssigneeRoche Diagnostics Operations, Inc.
InventorsManfred Watzele, Thomas Hoffmann, Cordula Nemetz, Dieter Heindl, Thomas Metzler, Wolfgang Mutter
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Abstract
The present invention concerns a method for improving the stability of linear short DNA towards exonucleases in cell-free in vitro transcription/translation systems using lysates containing exonucleases or in cellular systems containing exonucleases, wherein the stability of the linear short DNA is improved by adding unspecific linear DNA.
Claims16
A method for improving the stability of linear short DNA from degradation by exonucleases in cell-free in vitro transcription/translation systems that utilize lysates containing exonucleases or in cellular protein expression systems containing exonucleases, said method comprising adding unspecific linear DNA to the reaction in said systems.
A kit comprising one or several vessels which contain the following components:
- a lysate containing ribosomes prepared from prokaryotic or eukaryotic cells by a method for producing lysates for in vitro translation;
- one or more of the 20 naturally-occurring L-amino acids;
- one or more naturally-occurring or synthetic t-RNAs;
- one or more naturally-occurring nucleotides which can be unphosphorylated or be present as monophosphates, diphosphates or triphosphates;
- and linear unspecific DNA.
A reagent solution comprising:
- a lysate containing ribosomes prepared from prokaryotic or eukaryotic cells by a method for producing lysates for in vitro translation;
- one or more of the 20 naturally-occurring L-amino acids;
- one or more naturally-occurring or synthetic t-RNAs;
- one or more naturally-occurring nucleotides which can be unphosphorylated or be present as monophosphates, diphosphates or triphosphates;
- and linear unspecific DNA.
Dependencies are inferred from the claim text.