Biomate 3s uv visible spectrophotometer

Eggs laid by Fuyin, Fuyin-lre, and re were separately treated and incubated [13 ]. Every 24 h after incubation, the eggs were treated with 20% NaOH at 100°C for 3-5s, and then, eggs were blew with a dropper to separate the embryo from eggshell in clean water. The embryonic morphology of these eggs was observed under a microscope to determine the developmental stages of the embryo.
Every 24 h after incubation, 0.5 g of normal eggs laid by Fuyin as well as red eggs laid by Fuyin-lre and re was collected for total RNA extraction using the RNAiso Plus (TaKaRa) kit. The extracted RNA was digested by DNase I (TaKaRa) to remove DNA residues. The concentration and purity of total RNA was determined using a BioMate 3S UV-visible spectrophotometer (Thermo Fisher Scientific), and the quality of total RNA was identified using an Agilent bioanalyzer (Agilent Technologies 2100).The qualified RNA was used in DGE and qRT-PCR.
Normal eggs laid by Fuyin and the red eggs laid by Fuyin-lre and re were selected to extract their genomic DNA according to the literature [14 ]. Each qualified DNA sample was confirmed by the BioMate 3S UV-visible spectrophotometer before dilution to 100 ng/μL and storage at—20°C.

The amount of pcDNA3.1-SynORF5 plasmids loaded into PLGA NPs was determined as follows (Zhao et al., 2010 (link)). Lyophilized PLGA-SynORF5 NPs were immersed in 1 mL dichloromethane, to which 5 mL 0.1 M PBS (pH 7.4) were added. The mixture was stirred for 30 min followed by centrifugation at 4,000 rpm for 8 min. The supernatant was collected and the concentration of pcDNA3.1-SynORF5 was determined by measuring absorbance at 260 nm using Biomate 3S UV-Visible Spectrophotometer (Thermo Scientific, USA).
The amount of entrapped GP5 protein in NPs was determined as described previously (Corrigan and Li, 2009 (link)). Briefly, freeze-dried NPs were dispersed into 3 mL 0.1 M NaOH containing 5% (w/v) SDS. The suspension was incubated in a water bath at 60°C for 1 h. Following centrifugation at 10,000 rpm for 10 min, the concentration of the GP5 protein in the supernatant was measured by using RC DC^TM Protein Assay (Bio-Rad, USA) for micro-BCA analysis.
The EE of the NPs were calculated using this formula (Park et al., 2003 (link); Manca et al., 2008 (link)):
Where m is the mass of the pcDNA3.1-SynORF5 or GP5 loaded in PLGA NPs and C and V are the concentration and volume of the supernatant, respectively. m₀ is the initial amount of pcDNA3.1-SynORF5 or GP5.

NaYF₄:Yb,Tm nanocrystals and monodisperse silica-coated NaYF₄:Yb,Tm (NaYF4:Yb,Tm@silica, UCPs) were synthesized and characterized following the method described by Guo et al. (2010) [7] and Qian et al. (2009) [9] (link). The UCPs were modified using an amino group following the method described by Guo et al. (2011). DMNPE (Invitrogen) was used to cage siRNAs following the manufacturer's protocol, and UCPs/siRNA-DMNPE complexes were made as described previously by Guo et al. (2011) [14] (link). The ratio of UCPs to siRNA was determined by measuring the zeta potential using Nano-ZS ZEN 3600 (Malvern Instruments Ltd, UK) at 25°C. The UV absorption spectrum was gathered using BioMate 3S UV-visible spectrophotometer (Thermo Scientific) at 25°C.

For the pigment analysis, the leaves were collected from the sprl1 mutant and its wild type at the seedling stage and the heading stage, respectively. The pigments were extracted from 0.2 g of fresh leaves with 80% acetone in the dark at 4 °C for 48 h. The contents of chlorophyll (Chl) and carotenoids (Caro) were measured at wavelengths of 470 nm, 646 nm, and 663 nm using the BIOMATE 3S UV-Visible Spectrophotometer (Thermo scientific, Waltham, MA, USA) and were calculated according to the method of Lichtenthaler and Wellburn [52 (link)]:


For the Proto IX analysis, 0.1 g fresh leaves at the seedling stage were homogenized in 1 mL methanol/acetone/0.1 M NaOH (9:10:1, v/v/v), and the homogenates were centrifuged at 7197 g (Eppendorf 5430R; 7830 rpm) at 1 °C for 20 min. To oxidize the Protogen IX into Proto IX, 5 μL of 1 M acetic acid and 5μL of 2-butanone peroxide was added to 200 μL supernatant [28 (link)]. Then, the Proto IX was analyzed by HPLC on a C8 column (4.6 × 150 mm, 3.5μm; Waters) according to the method of Wang [53 (link)]. The elution profiles were detected by fluorescence, with excitation at 405 nm and emission at 625 nm [28 (link)]. The Proto IX was quantified by using the Proto IX standard (Frontier Scientific, Logan, UT, United States).