Invitrogen™

Retic Lysate IVT™ Kit

Catalog number:  AM1200

Related applications:

Protein Expression

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Description

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

Product Type: Kit
Quantity: 60 reactions
Expressed Protein Type: Water Soluble Protein
Expression System: Cell-Free
Expression Mechanism: Cell-Free Expression
Shipping Condition: Dry Ice
Product Line: Ambion™

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.