High Efficiency for Library Making

The Jackson Laboratory
Cancer biology
Post doc

Overall

Quality of Results

Ease-of-Optimization

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Company:

Invitrogen

Product Name:

Electro Max Stbl4 Cells

Catalog Number:

11635018

Stbl4 electro competent cells have high transformation efficiency which is useful for those who want to make a complex library directly from ligation mixture and culturing transformation mixture instead of plating them to a single colony.

Experimental Design and Results Summary

Application

To make a highly complex library preparation

Starting Material

Lentiviral plasmid

Protocol Overview

The ligation (concentrated from several ligation mixtures) was electroporated (2ul) in up to 20ul of Stabl4 cells @ 1200Kv, 25uF transformations were grown at 37C for 1hr. After that, the transformation was transferred to a 100ml culture overnight at 37C. The plasmid DNA was isolated by Maxi prep. The lenti library prepared from stabl4 was shown to be highly diverse compared to other competent cells.

Tips

Direct electroporation of ligation needs 90 min recovery time.

Results Summary

We made a highly complex library with diversity about 100000 different clones from plasmid size 15kb.

Additional Notes

None

Image Gallery

Summary

The Good

Good for complex library making

The Bad

None

The Bottom Line

Good electrocompetent cells to complex library making.

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