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The RiboPure™ kit samples showed the highest percentage of amplification and hence the largest amount of DNA contamination.
The required 200 ng RNA (10 ng/μl), was not achieved for the RNeasy® kit samples, or indeed for the RV2772 TRIzol® extracted sample.
For the CNA kit, samples were processed according to the manufacturer's protocol for 1-mL input volume with an elution volume of 50 μL.
After RNAse treatment (Epicentre RNAseH) and dscDNA purification (QIAGEN Qiaquick PCR purification kit), samples were labelled using Invitrogen BioPrime DNA labelling, and processed.
The number of total reads and viral reads obtained for the RNeasy® kit samples were lower in comparison to the TRIzol® extracted RNA, most likely due to the difference in total RNA available for these samples.
Where concentrations were above the detection range of the kit, samples were diluted according to the manufacturers' instructions to obtain a value within the detectable range, which was then multiplied by the dilution factor to obtain the final concentration.
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To prepare anti-pig IgGFc, 4 conjugate dilutions (1 1,000, 1 1,500, 1 2,000, 1 2,500) were titrated against 100 μl of kit negative and positive controls diluted 1 30 (10 μl kit control + 290 μl kit sample diluent).
Western blotting kits, sample buffer, molecular weight marker, nitrocellulose membrane, and filter paper for Western blotting were obtained from Bio-Rad LaBathtories (Corston, Bath, UK).
Using the newly designed primers and the AmpFl STR SGM Plus kit, 111 samples were examined.
DNA was again purified using MinElute PCR Purification kit and samples were sequenced using Illumina HiSeq 2500 system.
Taking out the kit of samples and the Ann Summers catalogue, Joanne was embarrassed, but her friends said: 'Don't worry about it.
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