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The washed pellet was resuspended in 3 ml extraction buffer and used for GBSS1 assay.
After three rounds of washing, sperm cells were resuspended in reaction buffer and used for incubation with the HPV16 capsids.
After 48 h, the transfected cells were extracted with lysis buffer and used for luminescence measurements with luminometer.
The recovered DNA was re-suspended in TE buffer and used for the PCR amplification as described [33], [34].
20 µl (2%) were added to SDS-containing sample buffer and used for SDS-PAGE (referred to as input).
Mouse embryonic fibroblasts were lysed with RIPA buffer and used for glycosidase analysis, SDS-PAGE and immunoblotting.
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Following the removal the supernatant, the resin was washed four times with ice-cold GST bind/wash buffer (Novagen) and used for the pull-down.
Eighty-five microfithe of the brain homogenate was mixed with 265 µL of lysis buffer (RLT) and used for RNA extraction.
Cells were then lysed on the plate using passive lysis buffer (Promega) and used for the Dual Luciferase Assay Kit following the manufacturer's guidelines (Promega).
The nuclear pellets were washed three times in RLN buffer (Qiagen) and used for isolation of nuclear RNA with the Qiagen RNeasy procedure for total RNA.
After this, cells were washed in Ca/Mg Phosphate buffered saline and used for confocal microscopy.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com