Quorum Sensing

to 1000 base pairs with most falling below 500 base pairs.

Figure3 shows four representative samples.

11. If pooling samples, bring each barcoded sample to 500 ng in
    4 μl of water. Then mix at an equal ratio by adding 1μl each to a
    tube and mix well. Consult the sequencing center of your
    choice for the desired final concentration and submit you
    sample(s) for sequencing on an Illumina Genome Analyzer
    (seeNote9).

4 Notes

1. The miRNeasy Mini Kit from Qiagen is recommended for
   RNA isolation. This kit allows for the isolation of total RNA,
   including microRNAs. If you plan to analyze microRNAs,
   please take this into account when mapping your sequenced
   reads and establishing cutoff parameters in data analysis. It is
   recommended that each condition be represented in at least
   two biological replicates.


2. Dabbing the tube on a paper towel or Kimwipe ensures that the
   RNAprotect Bacteria Reagent is removed. If too much reagent

Fig. 3 2% (wt/vol) agarose gel of four representative RNAseq library

preparations. Base pair size is indicated to theleftof the gel

188 Charlotte D. Majerczyk