Revision as of 09:53, 21 September 2011

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DNA Recovery Experiment

Introduction

Once the basic mechanism of cell lysing is confirmed, the next step is to show that DNA can be recovered from the supernatant.

The Experiment : The Experimental Setup for the Lysis-based DNA Recovery Experiment

The Results : The Results from the Lysis-based DNA Recovery Experiment

We grow two large cultures of cells. One contains cells that will lyse and release plasmids into the supernatant while the other has non-lysing, "normal" cells.

Adding IPTG to both flasks induces lysis in one set of cells but not in the others.

Thanks to qPCR, the supernatant harvested from the lysing culture reveals increased numbers of plasmids while the non-lysing culture exhibits significantly lower numbers of plasmids. A similar test involves transforming the supernatant (whose plasmid content is not known a priori) into cells and counting the number of resulting colonies. The plates containing transformations from the lysing supernatant have vastly superior number of colonies compared to the non-lysing supernatant transformations.

@@ Line 1: / Line 1: @@
 {{:Team:EPF-Lausanne/Templates/Header|title=DNA Recovery Experiment}}
-Having a lysis cassette driven by a T7 promoter is an important step towards being able to recover the DNA sequences of transcription factor and promoter mutants that have strong mutual affinities. To verify that basic lysing can be
+=== Introduction ===
-induced, we use the same IPTG experiment as for RFP, using both large samples for visual, qualitative confirmation and small samples in a platereader for quantitative, numerical confirmation.
+Once the basic mechanism of cell lysing is confirmed, the next step is to show that DNA can be recovered from the supernatant.
-Once the basic mechanism of cell lysing is confirmed, the next step is to show that DNA can be recovered from the supernatant. We grow two large cultures of cells. One contains cells that will lyse and release plasmids into the supernatant while the other has non-lysing, "normal" cells.
+* [[Team:EPF-Lausanne/Our Project/T7 promoter variants/dnarecov/exper| The Experiment ]]: The Experimental Setup for the Lysis-based DNA Recovery Experiment
+* [[Team:EPF-Lausanne/Our Project/T7 promoter variants/dnarecov/results| The Results ]]: The Results from the Lysis-based DNA Recovery Experiment
+ We grow two large cultures of cells. One contains cells that will lyse and release plasmids into the supernatant while the other has non-lysing, "normal" cells.
 [[File:broth_noiptg.png]]