The 2139 bp PXL1 coding region (PXL1∆KD) was amplified with the primers listed in Table S3 and then inserted into the expression vector pNL with NLuc at the C-terminus by using a recombination kit (Vazyme). The 825 bp SERK1 (SERK1∆KD) and the 927 bp BAK1 (BAK1∆KD) coding regions were amplified with the primers listed in Supplementary Table 3 . The PCR products of SERK1∆KD and BAK1∆KD were inserted into the expression vector pCL with CLuc at the C-terminus by recombination kit (Vazyme). The plasmids were introduced into Agrobacterium strain GV3101, and then single colonies were cultured in liquid YT medium. The OD values were measured with a UV/Vis spectrophotometer (Eppendorf). Then, resuspensions of well-cultured Agrobacterium containing the intended plasmid were mixed with 1:1 ratio as required and the different combinations were infiltrated into 4-week-old N. benthamiana leaves. After 2 days, 20 μm CLE19 was infiltrated into the expressing N. benthamiana leaves, with PBS as a negative control, 3–4 h before observation. Then, each infiltrated leaf was sprayed with 0.25 mM luciferin substrate and placed in the dark for 10 min, and LB985 NightShade (Berthhold Tech) with IndiGo software was used to detect the interaction signal.
Uv vis spectrophotometer
Manufactured by Eppendorf
Sourced in Germany
The UV/Vis spectrophotometer is a laboratory instrument used to measure the absorption or transmission of light in the ultraviolet and visible regions of the electromagnetic spectrum. It is designed to quantify the concentration of specific compounds in a sample by measuring the amount of light absorbed or transmitted at a particular wavelength.
Automatically generated - may contain errors
Lab products found in correlation
Recombination kit, by Vazyme (1 mentions)
Rnasimple total rna kit, by Tiangen Biotech (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
N60 nano spectrophotometer, by Implen (1 mentions)
Multidoc it digital imaging system, by Analytik Jena (1 mentions)
Advance spectrometer, by Bruker (1 mentions)
6538 q tof, by Agilent Technologies (1 mentions)
Fluoromax 4 fluorimeter, by Horiba (1 mentions)
Methanol, by HiMedia (1 mentions)
15 protocols using uv vis spectrophotometer
1
Protein-Protein Interaction Assay in N. benthamiana
2
Hydrolysis of p-NPP for Lipase Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The hydrolysis of p-NPP as a substrate used to estimate the activities of free and immobilized lipases as it is reported previously.30 (link) Briefly, a substrate solution which was a mixture of 9 volumes of solution B (0.4 mL of Triton X-100 and 0.1 g Arabic gum in 90 mL PBS; 50 mM, pH 7.5) and 1 volume of solution A (30 mg p-NPP in 10 mL isopropanol) was prepared. The reaction would take place at 37 °C by preincubating the substrate solution. For each activity assay, 1.8 mL of substrate solution and a specific amount of lipase or piece of membrane reacted for 2 minutes and released p-nitrophenol was measured at 405 nm in a UV/VIS spectrophotometer (Eppendorf) against the blank sample, which contains no enzyme. One unit of lipase activity (1 U) is defined as the amount of enzyme or biocatalyst, which liberates 1 μmol of p-nitrophenol per minute under the assay condition. Lipase activity and specific activity were determined as follows:
Relative activity was defined as the percentage of the maximum activity obtained in that series.
Relative activity was defined as the percentage of the maximum activity obtained in that series.
3
Microbial Cell Dry Weight Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Glucose and lactate concentrations were measured using Biosen C_line GP (EKF-diagnostic GmbH, Barleben, Germany). The OD was measured using a UV/Vis spectrophotometer (Eppendorf AG, Hamburg, Germany). An aliquot was diluted with reverse osmosis (RO) water to ensure the absorbance was <0.5. The optical density (OD) of the dilution was then measured three times at 600 nm (OD600). To determine the CDW, 10-mL aliquots of cells were collected by centrifugation (10,000 rcf, 10 min, 4 °C) and transferred to a dried, pre-weighed 2-mL centrifuge tube. The cells were then resuspended in RO water and centrifuged at 16,100 rcf for 2 min at 4 °C (Eppendorf AG). The pellets were washed, centrifuged twice as above and dried at 55 °C until a constant mass was obtained.
4
RNA Extraction from Ambrosia orientale
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from A. orientale roots, tubers, and leaves using the RNAsimple Total RNA kit (Tiangen, catalog number DP419). RNA samples were subjected to electrophoresis on 1.0% agarose gels and quantified by electronic digital imaging using the Syngene G:Box. RNA yield and purity were assessed using an Eppendorf UV-Vis spectrophotometer.
5
CTAB-based Genomic DNA Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Genomic DNA was extracted from young leaf tissue following the Cetyltrimethylammonium bromide (CTAB) based protocol of Murray and Thomson (1980) (link) with minor modifications. Briefly, the fresh leaves were homogenized in 20 ml extraction buffer (2% CTAB, 1.4 M NaCl, 100 mM Tris, 20 mM EDTA and 1.5% β-mercaptoethanol) at room temperature using a mortar and pestle, and the extract was incubated at 65 °C for 1 h. DNA was isolated by chloroform: isoamylalcohol (24:1, v/v), treated with RNase A (100 μg/ml, 30 min. at 37 °C) to avoid RNA contamination, precipitated with isopropanol and washed with ethanol. The quality of the DNA was checked by a UV–vis spectrophotometer (Eppendorf, Germany) by checking the A260/A280 ratio. The final concentration of DNA was adjusted to 50 ng/μl. All the DNA samples were stored at − 20 °C for genotyping.
6
Mammary Gland Total RNA Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was extracted from mammary gland tissue using Trizol Reagent (Invitrogen, Grand Island, NY, USA). The extracted RNA was dissolved in DEPC water, and then stored at −80°C for further analysis. The purity and concentration of RNA samples were determined by an ultraviolet-visible spectroscopy (UV/Vis) spectrophotometer (Eppendorf, Hamburg, Germany).
7
Quantifying Extracellular Matrix Composition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Twenty-four, 48 and 72 h WT and mutant strain cultures were centrifuged (1 min at 13 200 r.p.m.), after which the supernatant was discarded. Cell pellets were then resuspended in 1.5 M NaCl, as described previously [22 (link)]. The resuspended cells were then centrifuged once more, and the supernatant was collected to measure the concentration of DNA, proteins and saccharides. To quantify DNA in the isolated ECM, DNA was purified with the Promega SV Wizard DNA purification system (Promega, USA) according to the manufacturer’s instructions. The concentration of the purified DNA was measured using an Implen N60 nano-spectrophotometer (Implen, USA). Proteins in the isolated ECM fractions were quantified via the Bradford assay based on a bovine serum albumin standard curve. Protein concentration was measured at 590 nm on a UV/Vis spectrophotometer (Eppendorf, Germany). The total saccharide concentration in the ECMs was measured using the phenol–sulfuric acid method [22 (link)]. Briefly, 20 µl of 5 % phenol was added to 20 µl of the ECM fraction and mixed in the 96-well plate. Afterward, 100 µl of sulfuric acid was added, and the mixture was incubated for 10 min at room temperature. Saccharide concentration measurements were performed at 492 nm using a Spark (Tecan, Switzerland) instrument with glucose as a standard.
8
Metagenomic DNA Extraction and Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Equal volume (3 μL) of the extracted metagenomic DNA from all the samples for methods M1 to M6 were loaded in 0.8% agarose gel along with 3 μL of 1Kb DNA ladder (50 μg/mL) and the bands were visualized using UVP-Multidoc-It digital imaging system, CA, USA (Fig 1 , S2 Fig ). Purity and concentration of metagenomic DNA was determined by spectrophotometry analysis (UV-vis spectrophotometer, Eppendorf, NY). Concentration of the DNA (ng/μL) allowed calculation of Yield in μg per gram of soil = concentration of DNA (μg/μL)•volume used to suspend DNA (μL) /weight of soil (g) [24 ]. The absorbance A260/280 was used to determine protein contamination, and A340 as an indication of humic acid contamination.
9
Chlorophyll and Photosynthesis Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine total chlorophyll, 20 mg fresh leaves were taken from control and treated seedlings. Leaves were then subjected to crushing in 80% acetone followed by pigment extraction and centrifugation. Absorbance of the extract was taken at 663 and 646 nm by using Eppendorf UV-VIS Spectrophotometer. Total chlorophyll was measured according to the method of Lichtenthaler (1987) . For measurement of photosynthetic performance of seedlings, chlorophyll a fluorescence was observed in the dark-adapted (30 min) state of leaves with the help of hand-held leaf fluorometer (Fluor Pen FP 100, Photon System Instrument, Czech Republic). The fluorescence parameters including maximum photochemical efficiency of PS II (Fv/Fm), photochemical quenching (qP) and non-photochemical quenching (NPQ) were then measured.
10
Encapsulation Efficiency and Drug Loading of QD@MSN-NH2 Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Typically, 2 mg of QD@MSN-NH2 were dispersed in 1 ml EPI solution (1 mg/ml) and sonicated for 5 min. Then the mixture was stirred for 24 h at room temperature. Afterward, drug-loaded nanoparticles were separated by centrifugation (15,000 g for 10 min), and the supernatant containing free EPI was collected. The amount of free EPI was evaluated by absorbance of supernatant and equation of standard curve (Supplementary Fig. 7 ) at λ = 480 nm using a UV/Vis spectrophotometer (Eppendorf, Germany). To calculate the encapsulation efficiency (EE%) and drug loading capacity (LC%), the Eq. 1 and Eq. 2 were used:
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!