The Retic Lysate IVT™ Kit efficiently translates in vitro-synthesized transcripts, poly(A) RNA, and total RNA, including difficult-to-translate RNAs. The optimized lysate results in high protein yield, biological activity, and consistent performance. A report in Nucleic Acids Research (1) found that some commercial reticulocyte lysates had poor fidelity for initiation at the appropriate AUG initiator codon. The Retic Lysate IVT Kit was tested using Kozak's constructs and it demonstrated high fidelity for initiating at the proper AUG.
- Efficiently translates a wide variety of high molecular weight proteins and both capped and uncapped mRNAs
- Accommodates large volume of template mRNA
The lysate has been optimized for translation by the addition of an ATP-regenerating system, hemin, and calf liver tRNA. Included in the kit is an optimized Translation Mix (-Met or -Leu) for the translation of both capped and uncapped messages along with High and Low Salt buffers. These buffers can be used to maximize protein yield and activity from difficult-to-translate sequences.
1. Kozak, M. (1990) Evaluation of the fidelity of initiation of translation in reticulocyte lysates from commercial sources. Nucleic Acids Res 18:2828.
|Cell Free Protein Expression for Functional Analysis, Cell-Free (in vitro) Expression, Protein Expression, Proteins, Expression, Isolation and Analysis|
|Retic Lysate, 20X Translation Mix minus-Met, 20X Translation Mix minus-Leu, High Salt Mix minus-Met, Low Salt Mix minus-Met, Capped Xef-1 RNA, and Nuclease-Free water|
|Retic Lysate should be stored at -80°C. 20X Translation Mix minus-Met, 20X Translation Mix minus-Leu, High Salt Mix minus-Met, Low Salt Mix minus-Met, and Capped Xef-1 RNA should be stored below -70°C. Nuclease-Free water can be stored at any temperature.|