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Micropipette

Manufactured by Thermo Fisher Scientific

A micropipette is a laboratory instrument used to measure and transfer small volumes of liquids, typically in the range of microliters (μL) to milliliters (mL). It is designed to provide precise and accurate liquid handling for various applications in scientific research and analytical laboratories.

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3 protocols using micropipette

1

Evaluating Cell Recovery Rate in Foreskin Fibroblasts

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Example 5

Normal human neonatal foreskin skin fibroblasts (NHDF, KURABO INDUSTRIES LTD.) in the logarithmic growth phase (1440×104 cells) were prepared, suspended in the medium composition (48 mL) of Example C371 in Table 3, dispensed by 30×104 cells (1 mL) to a 24-well cell culture plate (manufactured by SUMITOMO BAKELITE CO., LTD.), and cultured under 37° C., 5% carbon dioxide gas conditions for 3 days. After culturing, the cell concentration of the cell suspension was measured by a cell counter (TC-20, BIO-RAD), the suspension was transferred to a 1.5 mL microtube and pipetted a given number of times using a micropipette (manufactured by Thermo Scientific, clip chip 1000 μL) set to suction/discharge volume 0.2 mL. Thereafter, centrifugation (300×g, 3 min) was performed and the supernatant (1.1 mL) was removed. The cells were resuspended by adding 10% fetal bovine serum-containing DMEM-LG (0.9 mL), and the amount of ATP contained in the cells was quantified by a plate reader (manufactured by Tecan Japan Co., Ltd.) and using CellTiter-Glo (Promega Corporation), and the cell recovery rate was calculated. The above test was performed 5 times each, and the mean thereof is shown in the Table.

TABLE 4
number of pipettings
35101520
cell recovery rate %44.269.970.668.375.9

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2

Embryo Development in Avian Eggs

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Egg shells were cleaned carefully by rubbing with a cotton swab damped in 70% alcohol. A small area of egg shell was softened with a small droplet of concentrated HCl-(Ittehad chemical limited) applied with the help of a micropipette-(Thermo Fisher) in the middle of the two tapered ends. Each egg was placed horizontally (window facing laterally) for 10 min, so that the embryo (germinal disc) should gradually migrate on the top of the yolk ball and avoided from any injury. A fresh disposable insulin syringe (1 mL) with stainless steel needle of 12 mm length was used to inject the relevant dose volume containing the required ingredients of the respective group through the soften egg shell area directly into the yolk ball in each egg. The dose was administered at zero day of incubation in all groups. Immediately after injection, the minute hole was properly sealed with melted wax to avoid contamination. Before incubation, weight of each egg was measured by electrical weight balance. The treated eggs of all three groups were placed in an incubator (Memmert) at 37 ± 0.5 ºC and 65% humidity for 14 days. During incubation all eggs were gently rolled manually, twice a day (12 hourly basis).
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3

Chitinase Production and Activity Assay

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Materials used in this research were chitin and semi-purified chitinase obtained from the previous study (Soedirga et al., 2019) , commercial powdered agar (Agarpac Swallow Globe), Potato Dextrose Broth (PDB) (Merck P6685 Specification: PRD.4.ZQ5.10000036227), MgSO4.7H2O (Merck CAS number:10034-99-8), (NH4)2SO4 (Merck CAS number 7783-20-2), phosphate buffer solution 0.1 M (Sigma Aldrich P5244 Specification: PRD.1.ZQ5.10000006487), distilled water (Amidis, 5 L gallon), 3-5-dinitrosalicylic acid (DNS) (Sigma Aldrich CAS number 609-99-4), Na-K, tartaric 4% (Merck CAS number 6381-59-5), N-acetylglucosamine standard (Sigma Aldrich A2278 CAS number 134451-94-8). While the equipment used were centrifuge (MPW-223e), centrifuge tube, glassware (Iwaki Pyrex), plastic syringe (Thomas Scientific), autoclave (Hiramaya), water bath (Cimarex), UV-VIS spectrophotometer (Thermo Scientific Genesys 10), quartz glass cuvette (Helma Analytics), pH meter (Methrohm 913), vortex (Cole-Parmer), analytical balance (Ohaus u-1800 ar 2140), filter paper (Sig, micropipette (Thermo fisher), and tip.
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