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White colonies on selective Luria Bertani (LB) agar plates were picked into 96-well blocks containing 1 ml LB broth plus kanamycin (50 µg ml−1) and grown overnight.
Transformants were picked into 50 ul of ultra-pure water and plasmid DNA liberated by heat lysing.
Plaques were picked into phage buffer and replated with bacteria for several rounds of infection to purify phage isolates.
Plaques were picked into 100 µl phage buffer (10 mM Tris-HCl, pH 7.5; 10 mM MgSO4; 68.5 mM NaCl; 1 mM CaCl2).
A total of ∼16,000 clones were picked into 96-well plates and replicated to 384-well plates for storage at −80°C.
Briefly, individual worms were picked into PCR tubes, lysed, and mtDNA copy number of individual nematodes was determined by quantitative real-time PCR (qRT-PCR,) using a reference sample of known copy number (quantified by serial dilution).
For identification, protein spots of interest were picked into a 96 well plate using the Ettan spot picker (GE Healthcare, Uppsala, Sweden) equipped with a 2 mm diameter picker head.
Plaques containing mixtures of deletion and wild-type DNA were picked into 100 µl buffer, and 10 µl of 10−3, 10−4 and 10−5 dilutions were plated with 300 µl M. smegmatis cells.
After confirming fluorescent and non-fluorescent colonies under a Leica MZ FL III stereomicroscope using Chroma filter (set #11003 BL/VIO) for each construct, approximately 30 transformed colonies were picked into 0.5 ml microcentifuge tubes containing 50 µl of Super Optimal Broth (SOC) media with Ampicillin.
Individual colonies were picked into 50μl of 0.01% Tritron-X.
Marker positive F1 animals were picked into 30 µl of M9 + gelatin in a 96 well.
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