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14 As a screen to identify materials with broad spectrum resistance to bacterial attachment, the polymer microarray were incubated with three different green fluorescent protein (GFP -labelled bacterial species, Pseudomonas aeruGFP -labelledStaphylococcus aureus 8325-4 and uropathogenic Escherichia coli O6:K15:H31 (UPEC) for 72 h.
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To visualise antibody binding, the microarray was incubated with 3′-diaminobenzide and counterstained with Mayer's Haematoxylin.
The microarray was incubated for 90 minutes at RT in the dark.
A protein microarray was incubated with a mixture of fluorescently labeled in vitro transcribed ASH1 mRNA and poly(A -selected totA -selectedm cells harvestotalt mRNAlog phase in YPD (fromre 1A).
The microarray was incubated overnight at 42°C in a hybridization cassette (Telechem International).
The hybridized microarray was incubated for 16 hrs at 42°C and then scanned with an Affymetrix 428 scanner (Affymetrix, USA).
For the detection the microarray was incubated with streptavidin HRP conjugate (R&D Systems) followed by dye precipitation reaction using TrueBlue™ (KPL, Gaithersburg, MD, USA).
First, the protein microarrays were incubated with a PBS blocking solution containing 1% BSA and 0.1% Tween20 for 1 hour at 23 °C to reduce non-specific binding.
Microarrays were incubated overnight (16 20 hours) at 50°C in a dark humidified chamber (Genetix).
For labeling, microarrays were incubated for 30 min with ExtrAvidin Peroxidase (Sigma) diluted 1∶1000 in BSA Peroxidase Stabilizer (BioFX).
Afterwards, the microarrays were incubated for 16 h in a wet chamber to ensure efficient covalent binding of the oligonucleotides to the surface.
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