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Subsequently, they were transformed into chemically-competent cells of DH5α and the inserts of purified plasmids were verified by direct DNA sequencing.
From these, 22 clones were purified, in vivo excised and the inserts of the plasmids were sequenced.
PCR products amplified with PfuUltra (Stratagene) were inserted into the pENTR/D-TOPO vector (Invitrogen), and the inserts of at least three independent clones for each clock gene were sequenced.
Eight randomly picked complements were analyzed, and the inserts of the plasmids revealed identical cDNA sequences for all plasmids.
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Amplification products were cloned in the pGEMT-Easy cloning vector (Promega Co, Madison, WI, USA) and the insert of at least 10 different colonies of each product were sequenced.
And the insert of the clones could not be entirely the full-length cDNA.
A mad tinkerer and devotee of duct tape, Salazar tried to alleviate the pain by constructing various pads and cutouts in the inserts of Ritzenhein's shoes.
Briefly, A549 and NCI-H1975 cells were seeded onto the inserts of Boyden chambers and cultured overnight.
The inserts of BEST310/BAC1-GL BEST310/BAC1-GLL were ando analyzed by iREX/BAC1-GLanalysis.
The inserts of these and all other plasmids were verified by sequencing.
Five of the six colonies analyzed contained an insert and the inserts were of different size.
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