Sentence examples for both the insert and from inspiring English sources
However, such methods usually require generating complementary single-stranded overhangs in both the insert and vector fragments, with or without RecA-mediation [8], [12], [14].
Although cytokine profiling results showed that PBMC from vaccine recipients respond to both the insert and the vector with similar cytokine production profiles, Ad5 stimulation of PBMC produced significantly higher amounts of IL-10 than Gag stimulation.
For synthetic biology applications involving high-complexity or multi-fragment cloning, sequence-dependent methods are generally inconvenient because they require unique and specific sites in both the insert and the vector in order to generate the initial plasmids [4] [6].
The final expression plasmid, pET K) MSP142 Camp.2, was prepared by digesting both the insert and pET K) vector DNA's with NdeI and NotI, the digested DNA's were gel purified, ligated and transformed into BL21-DE3.
In UDG-based methods, a ribonucleotide U replaces a T in the PCR primers used to add overlapping sequences to the insert and subsequent treatment with UDG enzyme generates single-stranded ends in both the insert and the vector for annealing.
For example, in LIC, overlapping sequences lacking a particular dNTP are added to the insert by PCR and complementary 12-nt single-stranded regions in both the insert and the vector are generated by T4 DNA polymerase treatment in the presence of that particular dNTP.
However, ligation-dependent cloning is often hampered by the requirement for multiple rounds of enzyme treatments and purification of both the inserts and vectors.
The AT1R antagonist valsartan (provided by Dominik N Müller, Max-Delbrück-Centre for Molecular Medicine, Berlin, Germany) or the ETAR antagonist sitaxsentan (provided by Pfizer, New York, NY, USA) was added as indicated into both the inserts and the corresponding wells.
After overnight incubation the medium was removed from both the inserts and external wells, and replaced in the external wells only by serum-free keratinocyte-defined medium consisting of KGM but without bovine pituitary extract and supplemented with 1.3 mM calcium (Sigma, Vienna, Austria), 10 μg/ml transferrin (Sigma), 50 μg/ml ascorbic acid (Sigma), and 0.1% bovine serum albumin (Sigma).
Subsequently, the circumference of both the insert periphery and the wear scars were digitised on calibrated digital images of the articular surface using published photogrammetry methods.
