A hybridisation buffer master mix was prepared for 3 library captures at room temperature containing 75 uL SureSelect hybridisation buffer #1, 3 uL SureSelect hybridisation buffer #2, 30 uL SureSelect hybridisation buffer #3 and 39 uL SureSelect hybridisation buffer #4.
Amplification reaction product was cleaned using Exonuclease I (New England Biolabs, Ipswich, MA, USA) and diluted 1 5 in Tris EDTA buffer, pH = 8. qRT-PCR mix was prepared using 2× SsoFast EvaGreen Supermix with low ROX (Biorad, Hercules, CA, USA).
For each sample, a 50 µl PCR mix was prepared containing 1×PCR buffer, 200 µM dNTP PurePeak DNA polymerase Mix Pierce Nucleic Acid Technologiess, Milwaukee, WI,), 0.5 µM of each primer (SGS, Köping, Sweden) and 2.5 U PfuUltra High-Fidelity DNA polymerase (Stratagene La Jolla, CA).
50 µL of each antibody mix was prepared in ELISA coating buffer (eBioscience, San Jose, CA) and coated on to ELISA plates (Maxisorp, Nunc).
A master mix was prepared containing 5× RT buffer, 40 U/μl RNAse inhibitor, 20U/μl Transcriptor reverse transcriptase, 10 mM each Deoxynucleotide Mix, 600 μM Random Hexamer Primer, RotaA.ext.
A PCR master mix was prepared using the following reactives: buffer (10×), bethain, dNTP 2.5 mM, Taq DNA polymerase rec (5U/ul Invitrogen), MgCl2 and H2O.
For primary inoculation, a DNA-plasmid mix was prepared in 20 μL using 50 mM potassium phosphate buffer, pH 7.0 (=mock buffer).
An RRL master mix was prepared by supplementing with the same buffer and ATP regeneration system used for yeast in vitro translation assays [46].
The sequencing mix was prepared with 63 μl nuclease-free water, 75 μl 2x running buffer, 8 μL DNA library, and 4 μL fuel mix.
Master mix was prepared by mixing the protein extract with 4X NuPAGE LDS sample buffer (Invitrogen, USA) and NuPAGE reducing agent (Invitrogen, USA).
