Tetracycline

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(Working concentration and Stock Solutions)
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*Using a plasmid with copy number 15-20, 20μg/ml tet has proved a successful selector.  The reduction in growth rate, described above was significant.
*Using a plasmid with copy number 15-20, 20μg/ml tet has proved a successful selector.  The reduction in growth rate, described above was significant.
*The RajBhandary lab uses 5μg/ml and see a minimal reduction in growth rate and never lose plasmid from their cells.
*The RajBhandary lab uses 5μg/ml and see a minimal reduction in growth rate and never lose plasmid from their cells.
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*Stock solution is 5 mg/ml in ethanol. Higher concentrations are not soluble at -20 C.  Solutions are light sensitive.
+
*Stock solution is 15 mg/ml in 50% ethanol.   Add 1 ml per liter of cooled broth or agar.  Solutions and plates are light sensitive.
==Plate Color Code==
==Plate Color Code==

Revision as of 14:38, 19 December 2005

Contents

Mode of Action

Tetracyclines (there are many) enter the cell by diffusion through OmpF porins. It inhibits growth by preventing codon-anticodon interactions during translation.

Resistance Mechanism

There are a number of resistance mechanisms, most require tetracycline antiporters. The most commonly used is the tetA(C) gene. The gene product of tetA(C) forms a pore in the inner membrane and exports one tet molecule for every proton imported. Molecular Cloning states that tet resistance has a number of adverse affects on the cell, chief among those being a reduction in growth rate and viability.

Working concentration and Stock Solutions

  • Using a plasmid with copy number 15-20, 20μg/ml tet has proved a successful selector. The reduction in growth rate, described above was significant.
  • The RajBhandary lab uses 5μg/ml and see a minimal reduction in growth rate and never lose plasmid from their cells.
  • Stock solution is 15 mg/ml in 50% ethanol. Add 1 ml per liter of cooled broth or agar. Solutions and plates are light sensitive.

Plate Color Code

Yellow stripe

Properties

  • Molecular Weight - 480.9g

References

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