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The isolates were checked for purity and preserved at −70°C for further studies.
After culture and selection of possible transformants, the isolates were checked for the correct insertion of the kanamycin resistance gene into the irp2 gene by DNA sequencing.
The isolates were checked to be free from African swine fever virus (ASFV) by using haemabsorption-inhibition test as well as ELISA.
The isolates were checked with PCR to assure the deletion and KanMX6 cassette was in the right place then back-crossed into a wt strain (h-, leura4-D18ra4-D18, ade6-M216).
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Purity of the isolates was checked by microscopic observation and further confirmed by denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA gene fragments (Teske et al. 1996).
Isolates were checked for the presence of 16 selected VF genes associated with extraintestinal infections and classified into 4 ECOR groups using PCR.
S. aureus isolates were checked for the presence of selected virulence genes: tsst (toxic shock syndrome toxin), sea, seb, sec, seg, seh, sei, sej (staphylococcal enterotoxins A, B, C, G, H, I, J), eta, etb (exfoliative toxins A and B), lukE (LukDE leucocidin), pvl (Panton-Valentine leucocidin, and hla (staphylococcal alpha haemolysin) using PCR with previously published primers [ 8- 10].
All isolates were checked for purity and cultivated on malt extract agar medium (Oxoid, Basingstoke, UK).
Second, we did not check MIC of daptomycin, and only 77.4% of MRSA isolates were checked for MIC of vancomycin.
The suitability of the primer pairs ASPF1CP ASPR3CP and ASPF2CP ASPR3CP for detection of 19 virus isolates was checked.
The virulence of the human isolate was checked in each experiment by infecting intratracheally BALB/c mice and recovering the yeast cells from their organs.
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