Faithful Amplification of Lenti Viral Vector

The Jackson Laboratory
Cancer biology
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Company:

Agilent Technologies

Product Name:

SURE electrocompetent cells

Catalog Number:

200227

We need to generate lentiviral vector for our expression studies on the HSC cells. We had problems with lentiviral vector amplification (unstable and more unwanted recombination) in the large culture of bacteria . So we choose these cells because this strain suppressed unwanted recombination.

Experimental Design and Results Summary

Application

Faithfull amplification of large constructs-lenti vector

Starting Material

Lenti plasmid/ ligation mixture

Protocol Overview

We electroporated 2ul ligation mixture in to 40ul of SURE competent cells @1700V, 200 Ohms with single pulse. We followed the other steps as recommended in the Manual.

Tips

14hr- optimum yield with less unwanted recombination

Results Summary

We do observe unwanted recombination with our 13 kb lenti viral vector however its comparatively lower compared to other vendor Electro competent cells.

Additional Notes

None

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Summary

The Good

Less unwanted recombination

The Bad

Much less yield

The Bottom Line

Better for faithfull amplication of lenti vector in a large volume of bacterial culture

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