Sample Preparation. Users are responsible for preparing their own samples; however, the Facility Manager must approve the prepared samples (see #6) and may disallow samples that are not stable and could damage the instrument. The Facility Manager will train Members in sample preparation and may provide assistance in special cases, but in general Members are responsible for their own samples.
Sample Preparation a. Laboratory Molded Specimens - Use cylindrical specimens that have been compacted using the gyratory compactor (AASHTO T 312). Specimen diameter must be 6 inches (150 mm) and a specimen height must be 4.5 inches +/- 0.2 inches (115 +/- 5 mm). Note 1 - Experience has shown that molded laboratory specimens of a known density usually result in a greater density (or lower air voids) after being trimmed. Therefore, it is recommended that the laboratory technician produce molded specimens with an air void level slightly higher than the targeted trimmed specimen. Determine the density of the final trimmed specimen in accordance with AASHTO T 166.
Sample Preparation. (1) Sample to be processed should correlate with stage and site to be treated.
Sample Preparation. ]” shall mean, individually and collectively, the major [...***...]
Sample Preparation. 1. Add 1 µL Activator to 100 µL patient sample or control. Mix and allow to sit at room temperature for 10 minutes.
Sample Preparation. The preparation of samples was conducted under stringent, diagnostic-quality conditions in the Emory University Anthropology Laboratories to ensure that no cross contamination of samples occurred. All immunologic testing was conducted at the laboratories of the Centers for Disease Control and Prevention’s Division of Parasitic Diseases (CDC, DPD) in Chamblee, Georgia. A number of techniques have been used to extract antigens from mummified remains for ELISA analysis. Prior to preparing the samples to be tested for this research, four methods were tested for antigen yield in sample processing. Skin samples from three individuals whose provenience has been lost were prepared following each sample preparation protocol. Protocol one followed Deelder and colleagues (Deelder et al., 1990; Xxxxxx et al., 1992). One hundred and fifty milligrams of sample tissue was homogenized with 1ml phosphate buffered saline (PBS) on ice in an all glass homogenizer and centrifuged at 16000 g for 35 minutes. The supernatant was mixed with an equal volume 14% trichloroacetic acid (TCA) solution to obtain a 7% solution and spun down at 16000 g for an additional 35 minutes. The supernatant was dialyzed against distilled water through an 8 kDa membrane for 24 hours at 4 C and desiccated. Samples were reconstituted in 250 μl of PBS + 0.05% Tween. Protocol two was identical to protocol one with the exception of the use of a steel homogenizer. In protocol three 150 mg of tissue was combined with 0.3 ml PBS in a polypropylene tube and subjected to three freeze/thaw cycles consisting of 10 sec in liquid nitrogen followed by 10 sec in boiling water. After three cycles, the samples were sonicated for 15 min in ultrasonic bath. Following sonication, samples underwent an additional freeze/thaw cycle. Suspensions were incubated for 24 h at 4 C warmed to 37 C for 30 min to solubilise remaining antigens and centrifuged at 14000 rpm for 20 minutes. An aliquot of the supernatant was used in the ELISA (Xxxxxxxx et al., 2008). Protocol four was identical to protocol three except that the length of submersion in liquid nitrogen and boiling water was increased to 30 sec per cycle. After preparation, samples from the three individuals prepared following each protocol were tested in a single assay. After comparison of the antigen yield from each method, it was concluded that protocol two was the most efficient means of sample preparation. Samples previously tested negative and positive by Xxxxxx and co...
Sample Preparation. 1. Prepare hemolysates of patient specimens and controls as instructed in the “Specimen Preparation” section.
Sample Preparation. Human calmodulin (CaM) was expressed and purified in Escherichia coli BL21(DE3) cells, as described F xxxxx://xxx.xxx/10.1021/jacs.2c02201 previously.9 Two site-directed mutations at A17C and A128C in the NTD and CTD domains of CaM, respectively, were introduced as sites of attachment for the nitroxide spin-labels.9 Full deuteration was achieved by growing the bacteria in deuterated minimal medium, with U-[12C/2H]-glucose as the sole carbon source. R1 nitroxide labeling was carried out with S-(1-oxyl-2,2,5,5-tetramethyl-2,5-dihydro-1H- pyrrol-3-yl)methyl methanesulfonothioate (MTSL; Toronto Research Chemicals), as described previously.9 The purity of A17C-R1/ A128C-R1 nitroxide-labeled calmodulin was verified by mass spectrometry. Solutions for DEER EPR comprised 50 μM CaM (A17C-R1/ A128-R1), 8 mM CaCl2, 100 mM NaCl, 25 mM d-HEPES pH 6.4, and 20% (v/v) d8-glycerol, placed in 1.0 mm inner diameter (1.2 mm outer diameter) quarts EPR tubes (VitoCom). Freezing of the DEER samples was carried out using three different approaches: slow rate freezing (∼40 s) by placing the EPR tube in a −80 °C freezer;43 intermediate rate freezing (∼1.5 s) by directly placing the EPR tube in liquid N2;43 and rapid freeze-quenching (∼0.5 ms) by spraying a high- speed jet onto a spinning copper plate cooled to 77 K, as described previously.18,45 Q-Band DEER. Pulsed EPR data were collected at Q-band (33.8 GHz) and 50 K on a Bruker E-580 spectrometer equipped with a 150 W traveling-wave tube amplifier, a model ER5107D2 resonator, and a cryofree cooling unit, as described previously.52 DEER experiments were acquired using a conventional four-pulse sequence (Figure S1).53 The observer and XXXXX pump pulses were separated by ca. 90 MHz, with the observer π/2 and π pulses set to 12 and 24 ns, respectively, and the XXXXX π pulse to 10 ns. The pump frequency was centered at the Q-band nitroxide spectrum located at +40 MHz from the center of the resonator frequency. The τ1 value of 350 ns for the first echo period time was incremented eight times in 16 ns steps to average 2H modulation; the position of the XXXXX pump pulse was incremented in steps of Δt = 10 ns. The bandwidth of the overcoupled resonator was 120 MHz. All DEER echo curves were acquired for tmax = 7.5 μs, with the exception of the DEER echo curve for τ2 < 7.5 μs, where tmax was set to the value of τ2. DEER data were recorded with values of the dipolar evolution time T (=2τ2) set to 10, 15, 20, 25, 30, and 35 μs. Measurement t...
Sample Preparation. If testing 21 to 40 samples, remove two disposable Applicator Blade Assemblies from the packaging. If testing fewer samples, remove only one Applicator Blade Assembly from the packaging. Remove the protective guards from the blades by gently bending the protective piece back and forth until it breaks free.
Sample Preparation. Annex 6 A surface treated sample, taken from the production, shall be damaged by cross engraving (ISO 2409:2007) and stone impact (ISO 20567-1:2005) to represent ...”
