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The influence of porosity of carbon inserts on the cell performance is also studied by varying the porosity of carbon inserts in the range of 60 70%, 70 80%, 80 90% respectively.
After size-selection on agarose gel to recover fragments with inserts in the range of 100 200 bp, the library was amplified by PCR for 18 cycles.
Fragments were purified on agarose gel to recover fragments with inserts in the range of 160 240 base pairs.
A total of 29,300 high-quality capillary reads were produced mostly from two subclone libraries (libraries with inserts in the range 2 6 kb using the vector pOTW12 and libraries with inserts in the range 5 12 kb using the vector pMAQ1Sac_ BstXI).
Restriction digestion analysis was carried out to determine the length of the insert of the positive clones and found insert length in the range of 38 to 41 Kb (Additional file 1: Figure S1D).
The indexed paired‐end libraries had an insert size in the range of 150 300 base pairs.
The colonies which contained insert sizes in the range of 600-1.2 kb were selected for sequencing.
Sequencing of these clones indicated the insert size in the range of 50 bp to 792 bp with an average size of 309 bp.
The sizes of the assembled inserts were in the range 21.2 37.9 kb, with G + C contents revolving around 58.8 (±6.4) %.
The apparent permeability (Papp) to [C]mannitol measured across the same inserts was in the range 0.1 2.6×10−5 cm/s (Fig. 10), and showed an inverse relation to the TEER.
To subsequently merge contigs into scaffolds, it is necessary to generate additional libraries with long-insert sizes in the range of 3 40 kb (Fig. 2).
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