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TAG fraction was eluted from Silica Gel by adding three times 1 mL chloroform and transferred to a previously weighted glass flask.
The obtained pellet was resuspended in distilled water and further precipitated by adding three times volume of cold ethanol.
After calculating the mean and the standard deviation, the LOD (signal-to-noise ratio (S/N) of 3) was determined by adding three times the standard deviation and the LOQ (S/N of 10) by adding ten times the standard deviation to the mean.
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Labelling was performed by incubating in a 37 °C incubator for 20 min and stopped by adding five times the original staining volume of culture medium (containing ≥1% protein).
The NHS biotin was then precipitated by adding ten times volume of cold diethyl ether and was again washed with ether.
Proteins in the soluble fraction were precipitated by adding ten times the volume of 10% cold trichloroacetic acid.
After 10 min of incubation at 37°C the staining was quenched by adding five times the volume of ice-cold PBS and excess stain was washed off by repeated washes in PBS.
The SDS concentration was reduced gradually to 0.10% by adding five times of 50 ul (1 x DpnII digestion buffer or NEB buffer 4 for multiple enzymes) with mixing.
First, Genome Estimation size 1 (Ge1) was calculated to account for chromosome ends by adding two times the average spacing of framework markers to the length of each linkage group [ 52].
The genomic DNA (gDNA) was precipitated by adding two times the volume of the collected supernatant of 100% ice-cold ethanol and 1/10 of the volume of the collected supernatant of a 3 M sodium acetate solution and incubating for 30 min at −20 °C.
(2) Then, we modify the structure of PIT by adding two domains: receive time and tolerance time, which are used to record the Interests' arrive time and tolerance time respectively.
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CEO of Professional Science Editing for Scientists @ prosciediting.com