OriC/ter ratio determination

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Revision as of 04:27, 27 February 2009 by Torsten Waldminghaus (Talk | contribs)
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Contents

Curators

Torsten Waldminghaus

Anyone should feel free to add themselves as a curator for this consensus protocol. You do not need to be a curator in order to contribute. The OpenWetWare community is currently discussing the idea of protocol curators. Please contribute.

Abstract

This protocol describes how one can determine the ration of the origin of termination oriC and the terminus of replication ter in Escherichia coli. That is for example important for cell cycle analysis. To get the ratio one could use Southern Blot and quantification of the bands. However, this protocol describes the use of qPCR which is a bit faster.

Materials

List everything necessary to perform the protocol here. Include all information about suppliers, ordering details, etc. Links to the suppliers' page on that material are also appropriate and encouraged. Please be aware that users of this protocol may not be working in the same country as you.

Reagents

Biological resources e.g. cell lines, buffers (link to a method for making them), enzymes, chemicals, kits, etc.

Equipment

Any equipment used to perform the protocol (link to a method for using them).

Procedure

A step by step guide to the experimental procedure.

If you find it helpfull you could use the following icons to highlight important parts:

Image:Difficult step.png icon to highlight difficult steps

Image:Critical step.png warn about critical steps

Image:Time required.png help people plan how long steps will take

Image:Pause point.png where protocol can be interrupted

Image:Optional step.png steps that can be omitted or included

Critical steps

Referenced from the main protocol, a more thorough explanation of particularly important steps in the protocol.


Notes

Referenced from the main protocol, any comments about the protocol should be made here; i.e. how it was developed. Any comments added should be signed (by adding *'''~~~~''': in front) and explained. Links to FAQs/tips provided by other sources such as the manufacturer or other websites would be best made here.
Anecdotal observations that might be of use to others can also be posted here; e.g. 'my cells were still floating'.


Discussion

You can discuss this protocol.

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