Quorum Sensing

20. Agarose.


21. Electrophoresis apparatus.


22. PCR machine and tubes.


23. A primer set (Forward and Reverse) for a gene in your
    organism.


24. PCR reagents.

2.2 RNAseq Library
Construction

1. Sterile, RNase-free tips.


2. Microfuge and microfuge tubes.


3. 200 U/μl SuperScript®III Reverse Transcriptase (Invitrogen).


4. 10 U/μl DNA Polymerase I (Invitrogen).


5. 10 U/μlE. coliDNA Ligase (Invitrogen).


6. 2 U/μl RNase H (Invitrogen).


7. Invitrogen Second-Strand Buffer.


8. 0.2 M ethylenediaminetetraacetic acid (EDTA).


9. QIAquick PCR Purification Kit (Qiagen).


10. Quick Blunting™(New England Biolabs).


11. Quick Ligation™(New England Biolabs).


12. Deoxynucleotide Solution Mix (New England Biolabs).


13. 5 U/μl Taq DNA Polymerase with Standard Taq Buffer (New
    England Biolabs).


14. Ethanol.


15. Agencourt AMPure XP Kit (Fisher Scientific).


16. Magnetic bead stand (such as a MagneSphere, Promega).


17. Setup for DNA electrophoresis: casting tray, comb, agarose,
    running buffer, loading dye, ethidium bromide (EtBr).


18. PCR Primers (HPLC purified):
    (a) Forward (Fwd) Primer (AATGATACGGCGACCACCGA)
    (b) Reverse (Rev) Primer (CAAGCAGAAGACGGCATACG)


19. Adapters (NON-Barcoded) (HPLC purified):
    (a) FwdAD (AGATCGGAAGAGCGTAATGATACGGCGA
    CCACCGACACTCTTTCCCTACACGACGCTCTTCC
    GATCT)
    (b) RevAD (AGATCGGAAGAGCTCCAAGCAGAAGAC
    GGCATACGAGCTCTTCCGATCT)


20. Adapters (Barcoded) (HPLC purified):
    (a) FwdAD-AGT (ACTAGATCGGAAGAGCGTAATGATA
    CGGCGACCACCGACACTCTTTCCCTACACGACGC
    TCTTCCGATCTAGT)

RNAseq of QS Regulons 181