Rabbit Reticulocyte Lysate System for DNA

Rabbit reticulocyte lysate (RRL) protein synthesis technology is a robust eukaryotic tool for the expression of functional mammalian proteins in a simple procedure. Both DNA and mRNA are acceptable for the RRL system as the template.

Over the years, Creative BioMart has been committed to the expression of recombinant proteins in different biological systems. We have developed a system for clients who intend to provide linear or plasmid DNA as a template.

Technology Overview

The RRL protein synthesis system we offer is quite simple and efficient. It couples the process of template transcription and translation together. The whole system consists of:

Rabbit reticulocyte lysate mixed with all reagents needed for the coupled transcription/translation. (like RNA polymerases, amino acid mixture minus methionine and RNase ribonuclease inhibitor)
A DNA template that contains either T7, T3 or SP6 promoter

Standard in vitro RRL Process		Transcription/Translation-Coupled RRL Process
Linear DNA template by restriction digestion	1 hour	Add DNA template and perform coupled transcription and translation	1.5 hours at 30°C
Extract and precipitate DNA	0.5 hour	Separate translation products by SDS-PAGE	1.5 hours
Perform in vitro transcription	1 hour	Fix and dry gel	2 hours
Remove template with DNase	1 hour	Autoradiography	3 hours, -70°C
Extract and precipitate RNA	0.5 hour
Perform in vitro translation with LLR	1.5 hours at 30°C
Separate translation products by SDS-PAGE	1.5 hours
Fix and dry gel	2 hours
Auto radiography	3 hours, -70°C
Time required: 12 hours		Time required: 8 hours

A variety of proteins ranging in size 10–150kDa can be expressed through the system. The template will be added to the lysate mixture directly and incubated for 60–90 minutes at 30°C.

Requirements of the Provided Template

Rabbit Reticulocyte Lysate System for DNA

The TEMPLATE is the only thing that needs to be provided by the client. Here are some corresponding suggestions that you may need when designing your template.

The provided template should be linear or plasmid DNA.
The intended protein size should be 10–150kDa.
Please make sure that the target sequence should be located downstream of the T7, T3 or SP6 promoter.
Clients can also authorize us to design or synthesize the template. Just let us know your requirements.

Related Service

If you want to provide mRNA as the template for your RRL protein expression, please go to the Rabbit Reticulocyte Lysate System for mRNA page.

Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP.

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