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If the buffer was not exchanged completely (by reducing the number of centrifugations), remnants of proteinase K and the buffer interacted with Phi29 polymerase and hampered the amplification.
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Words are not exchanged.
The buffer was changed daily as to ensure the buffer was not saturated with either SNAP nor silicone oil.
(sml-buffer): Query if the current buffer is not a *sml*.
The buffer was then exchanged with distilled water and dialysis was continued for 24 h.
To confirm H-bonds formed in the secondary structures, the H-D exchange 1H-15N HSQC experiments were also performed after the 15N-labeled hβ4-loopsamplein H2O NMR buffer was exchanged into D2O NMR buffer through dialysis.
Each active fraction was concentrated 10 times through centrifugation via MCWO 10 Vivaspin 500 (Sartorius) and the buffer was exchanged to 25 mM potassium phosphate buffer, pH 6.0.
Finally, pure and homogenous fractions were collected and buffer was exchanged with a buffer containing 20 mM KPi (pH 7.6), 100 mM NaCl and 0.15% DDM.
The protein buffer was exchanged with 10 mM phosphate buffer, pH 7.4, containing 150 mM NaCl, using the centrifugal ultrafiltration filter.
Thymic fragments were pelleted by centrifugation at 300× g and the digestion buffer was exchanged.
Afterwards, the buffer was exchanged to a 50 mM glycine buffer at pH 10 to stop the protease activity and to prepare for cation exchange chromatography.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com