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We are grateful to Dr. Guillaume Chanfreau for detailed and insightful comments on the manuscript, Dr. Christos Delidakis for the generous donation of the Groucho monoclonal antibody, Dr. Richard Jones for the wing disc ChIP protocol and to Dr. Larry Marsh for the histone deacetylase inhibitor fly media preparation protocol.
Herein, we report both the source of media components and the media preparation protocol can impact yeast one-hybrid and two-hybrid results.
Using both yeast one-hybrid and two-hybrid experiments, we have identified potential sources of these problems: the media preparation protocol and the source of the yeast nitrogen base may not only impact signal range but also effect whether a result appears positive or negative.
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The bio-relevant dissolution media, FaSSIF phosphate buffer, FeSSIF acetate buffer, FaSSIF, and FeSSIF were prepared according to Phares SIF Powder Preparation Protocol.
Plasmid DNA was prepared using the Qiagen plasmid preparation protocol.
Pooled and purified PCR product was used to prepare the DNA library following the Illumina TruSeq DNA library preparation protocol.
For C. micaceus unamplified libraries were generated using the Pacific Biosciences standard template preparation protocol.
This sample preparation protocol has been described previously for SEM of biofilms47.
We have automated a complex library preparation protocol for transcriptome sequencing.
Then, the same library preparation protocol used for viral libraries was applied without significant modifications.
(a) Shows an example of electropherogram obtained following the metagenomic library preparation protocol described in paragraph 4.1.
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