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RNA was extracted using the RNeasy system (Qiagen).
RNA was isolated from whole animals using the RNeasy system with on-column DNase treatment (Qiagen) according to manufacturers instructions.
The lungs were mechanically homogenized, then total RNA was isolated from lung homogenates using the RNeasy system (Qiagen), and cRNA was synthesized and amplified from equal masses of total RNA using the Ilumina TotalPrep RNA amplification kit (Ambion).
The effect of these clones or pharmacological agents on E7 expression were tested in an RT-PCR assay as described below and none of them had any effect on E7 message (Figure S1) Total RNA was extracted from cells using the RNeasy system (Qiagen, Valencia, CA) and treated with RNAase-free Dnase (Qiagen) for 15 min at room temperature.
Isolated AMs were lysed, and total RNA was extracted using the RNeasy system (Qiagen, Alameda, CA).
Total RNA was extracted using the RNeasy system (Qiagen, Valencia, CA, USA).
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Both the positively and the negatively selected cells using the IFNγ secretion assay, as well as unselected PBMCs were lysed in RLT® buffer (Qiagen, Hilden, Germany), and total RNA was gained using the Rneasy® system (Qiagen) according to the manufacturer's instructions including DNAse treatment.
Both OC and cultivated cells were lysed in 600 μl lysis buffer with 6 μl mercaptoethanol, by using the RNeasy® system and reverse transcription was done with Omniscript™ RT Kit (all Qiagen, Hilden, Germany), in accordance with the manufacturer's instructions.
Total RNA was extracted using the RNeasy purification system (Qiagen, Manchester, UK).
RNA was isolated from hNPCs or differentiated cells using the RNeasy mini system (Qiagen, Hilden, Germany).
RNA was isolated using the RNeasy Kit system (Qiagen), performing a DNAse digestion according to the manufacturer's protocol.
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