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The Prepare Protein protocol package in Discovery Studio suite was used for inserting missing atoms in incomplete residues, modeling missing loop regions [ 82], deleting alternate conformations (disorder), standardizing atom names, and protonating titratable residues using predicted pKs [ 83].
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The DNA coding for crtW was inserted into the Sfu I and EcoR I restriction sites of plasmid pGAPZαA, and the same restriction sites were used for inserting crtZ DNA into pGAPZαA*.
The diagonal matrix (after decomposition on SVD) S is used for inserting the watermark after decomposition on wavelet transform.
Cultures of transformed E. coli were grown over-night in 100 ul LB media containing 100 ug/ml ampicillin, the cells were centrifuged and suspended in sterile distilled water, heat denatured for 10 mins at 95°C, 1 ul of supernatant was used for insert amplification with M13 forward and reverse primers.
In this study vitamin E blended ultra-high molecular weight polyethylene (VE-UHMWPE), which is one of the new generation material, was used for acetabular inserts.
The KAPA HiFi Ready Mix system was used for amplifying inserts, using the following thermal profile: initial denaturation at 95 °C for 5 min, followed by 35 cycles each of 98 °C for 30 s, the corresponding primer's Tm (Table 1) for 20 s, 72 °C for 2 min and a final extension of 72 °C for 5 min.
Subsequent alignment using Illumina's CASAVA pipeline (version 1.8.2) to the P. taeda chloroplast genome and available genomic sequence was used for determining insert-size statistics.
An 18-gauge needle (150 mm long) was used for lipoinjection and inserted subcutaneously at one of four points indicated in Figs. 2A and B. The operator took care to insert and place the needle horizontally (parallel to the body) to avoid damaging the pleura and causing a pneumothorax.
A thermal asymmetric interlaced PCR (TAIL-PCR) protocol [40] was used for cloning genomic DNA flanking inserted T-DNA from the insertion mutants.
Another catheter, which was used for injection of ink, was inserted and placed in the abdominal aorta from the right femoral artery.
For patients who had more than one ICU admission during their hospital stay, the ICU admission during which the tracheostomy was inserted was used for data analysis.
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