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For real time RT-PCR, cDNA synthesis was carried out using the AffinityScript qPCR cDNA Synthesis Kit (Stratagene) according to the supplier recommendations, adding 200 ng total RNA.
For example, an engineer was asked to "consider" being more open to supplier recommendations and to visit the supplier for a day.
One Twitter user said: "As Ecotricity have billed me extra to foolishly fund the Labour party, I shall now be needing a new energy supplier – recommendations?" At an event in Manchester on Tuesday night the Green party leader, Natalie Bennett, was expected to attack Labour for not supporting a blanket ban on fracking.
In Ethiopia, common bleach is however sold without supplier recommendations for use in food, which increases the risk of misuse.
Genomic DNA was eliminated by on-column DNase digestion according to supplier recommendations.
NHEK at exponential growth phase were treated twice by 50 µM H2O2 at 48 hrs interval and then electroporated with the mRFP-GFP-LC3 [18] or the mRFP-GFP control vector using the Neon transfection system (Invitrogen) according to supplier recommendations.
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Freelancer platforms, which are mostly for short-term projects, and supplier recommendation platforms, which help match you to an agency but are "ineffective when you need to find the right team".
Recombinant proteins were expressed in E. coli BL21 (codon +) strain, grown in 200 ml of auto induction media terrific broth (Formedium™), and purified on Glutathione Sepharose 4B beads essentially following the supplier recommendation (GE Healthcare).
For live staining imaging, 10 μM of carboxyfluorescein succinimidyl ester (CFSE green, excitation 492 nm/emission 517 nm, Molecular Probes) was added 30 min before microscopic analysis following the supplier recommendation.
As previously described16, all cell lines were cultured in optimal medium according to the suppliers' recommendations.
The restriction enzyme digestions, ligations and transformations were performed as the suppliers' recommendations.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com