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. Advantages of the Retic Lysate IVT™ Kit:
• Efficiently translates a wide variety of high molecular weight proteins and both capped and uncapped mRNAs • Accommodates large volume of template mRNA
How it works 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. 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.
Reference 1. Kozak, M. (1990) Evaluation of the fidelity of initiation of translation in reticulocyte lysates from commercial sources. Nucleic Acids Res 18:2828.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Format
Kit
Expressed Protein Type
Water Soluble Protein
Expression Mechanism
Cell-Free Expression
Expression System
Cell-Free
For Use With (Application)
In Vitro Transcription
No. of Reactions
60 Reactions
Product Line
Ambion™
Product Type
Retic Lysate
Quantity
60 Reactions
Shipping Condition
Dry Ice
Contents & Storage
• 5 × 200 μl (store at -80°C) • 2 × 1.75 ml Nuclease-free Water (store at any temperature) • 100 μl 20X Translation Mix minus-Met • 100 μl 20X Translation Mix minus-Leu • 100 μl High Salt Mix minus-Met • 100 μl Low Salt Mix minus-Met • 50 μl Capped Xef-1 RNA (50 μg/ml)
Unless otherwise stated, store reagents below -70°C. Do not store cell-free translation systems in the liquid phase of a liquid nitrogen storage system.