The gel was viewed using a Kodak Gel Logic 100 Imaging System (Kodak, Rochester, NY, USA). The magnitude of the expression of each gene was assessed by band densitometry using Kodak Molecular Imaging Software. The amounts of the c-fos and c-jun PCR products were normalized to those of β-actin. The expression levels of the c-jun and c-fos genes were presented as the means ± standard deviation of the average relative band densities.
Gel logic 100 imaging system
Manufactured by Kodak
Sourced in United States
The Gel Logic 100 Imaging System is a compact and versatile lab equipment designed for the visualization and documentation of DNA, RNA, and protein gels. The system incorporates a high-resolution camera, adjustable lighting, and image analysis software to capture and analyze gel electrophoresis results.
Automatically generated - may contain errors
Lab products found in correlation
Molecular imaging software, by Kodak (1 mentions)
M mlv rt, by Thermo Fisher Scientific (1 mentions)
Hipi pcr premix, by Elpis Biotech (1 mentions)
Xp thermal cycler, by Bioer (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Enhanced chemiluminescence kit, by Cytiva (1 mentions)
Sirt1, by Abcam (1 mentions)
α tubulin, by Santa Cruz Biotechnology (1 mentions)
Ultrapure agarose, by Thermo Fisher Scientific (1 mentions)
Zetasizer nano zs zen3600, by Malvern Panalytical (1 mentions)
Laemmli sample buffer 4, by Bio-Rad (1 mentions)
Mini protean 3, by Bio-Rad (1 mentions)
Powerpac basic, by Bio-Rad (1 mentions)
Coomassie brilliant blue, by Bio-Rad (1 mentions)
Developing buffer, by Thermo Fisher Scientific (1 mentions)
Tris glycine sds running buffer, by Thermo Fisher Scientific (1 mentions)
Tris glycine sds native sample buffer, by Thermo Fisher Scientific (1 mentions)
Novex zymogram gelatin gel, by Thermo Fisher Scientific (1 mentions)
Zymogram renaturing buffer, by Thermo Fisher Scientific (1 mentions)
Coomassie blue r 250, by Bio-Rad (1 mentions)
17 protocols using gel logic 100 imaging system
1
Quantifying c-fos and c-jun Expression
2
RT-PCR Analysis of SRG3 Gene Expression
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Total mRNAs were isolated from CD4+ cells using the Easy Red reagent (Intron, Korea) and were reverse-transcribed into cDNAs with oligo(dT) primers and M-MLV RT (Invitrogen Life Technologies, USA), according to the manufacturers’ instructions. PCR amplification was performed according to the recommended protocols for the PCR Amplification Kit; for each PCR amplification, 1 μg of cDNA, 5 pmol of each primer, and HiPi PCR Premix (ELPis Biotech, Korea) were included in a 20 μl reaction volume. PCR was conducted in an XP Thermal Cycler (Bioer Technology, Hangzhou, China) with the following thermal cycling parameters: 35 cycles of 94°C for 5 min, 94°C for 45 sec, 57°C for 45 sec, 72°C for 1 min, and 72°C for 5 min for SRG3 and 30 cycles of 94°C for 5 min, 94°C for 30 sec, 54°C for 30 sec, 72°C for 35 sec, and 72°C for 5 min for β-actin. Equal amounts of RT-PCR products were electrophoresed on a 1.5% agarose gel and stained with ethidium bromide. The intensity of the resulting bands was measured with a Gel Logic 100 Imaging System (Kodak, NY, USA) and analyzed using the Image J software package (National Institutes of Health, USA). The sequences of the PCR primers were as follow: β-actin forward, 5’-GTA TGG AAT CCT GTG GCA TC-3’ and β-actin reverse, 5’-AAG CAC TTG CGG TGC ACG AT-3’; SRG3 forward, 5’-GAC TAG ACC AAA CAT CTA CCT C-3’ and SRG3 reverse, 5’-GTC AAC TGA GCG ACT GGA TC-3’.
3
Western and Lectin Blotting of Kv3.1b Glycoproteins
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Total cell membrane and whole cell lysate samples for western and lectin blotting, respectively, were electrophoresed using 10% SDS gels for 1.7 h at 20 mA [16 (link), 18 (link)]. In brief, separated proteins were blotted to PVDF membranes (Millipore, MA, USA) for 2.5 h at 250 mAmps. Blotted membranes were incubated and developed. Unglycosylated and glycosylated Kv3.1b proteins were detected using the mouse anti-Kv3.1b antibody (Neuromab, CA, USA). Biotin-conjugated Phaseolus vulgaris Erythoagglutinin (E-PHA) or Phaseolus vulgaris Leucoagglutinin (L-PHA) (Vector Laboratories, CA, USA) was employed to probe membranes containing separated glycosylated proteins. Images were acquired using Kodak gel logic 100 imaging system.
4
Molecular Characterization of Bacterial Symbionts
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Internal transcribed spacer (ITS) 16–23S rRNA was PCR amplified using primers ITS322 and ITS340 [24 (link)]. An inner region of the 16S ribosomal RNA (rRNA) sequence was amplified using the universal primers 27f and 1525r [25 (link)]. The housekeeping genes atpD, glnII, and recA were amplified using the primers atpD273f-atpD771r [26 (link)], glnII12F-glnII689R, and recA41F-recA640R [27 (link)], respectively. The symbiotic genes nodA, nifH, and nodC were amplified using the primers nodA3F-nodA4R [28 (link)], nifHI–nifHF, and nodCF, nodCFn and nodCI, respectively [29 (link)]. The conditions used for PCR amplification were those described by the authors, with a modification for nodA as described by Tartaglia et al. [7 (link)]. PCR products were run at 100 V in Tris-acetate buffer pH 8.2 in 1.2% agarose gels, with 1-kb ladder (Maestro) and stained with SYBR™ Safe DNA Gel Stain. Gel images were captured using the Kodak Gel Logic 100 Imaging System. The PCR products were sequenced by Macrogen (Macrogen, Inc., Seoul, Republic of Korea).
5
Western Blot Analysis of Protein Targets
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Total proteins containing 30–80 μg were separated on 8% SDS‐polyacrylamide mini gels and transferred to nitrocellulose membranes. After blocking, the blots were incubated with antibodies for NF-κB p65, iNOS, lamin A/C, α-tubulin (Santa Cruz Biotechnology, Dallas, TX, USA), and SIRT1 (Abcam) in PBS or Tween 20 for 1 h, followed by two washes in PBS or Tween 20, and subsequently incubated with horseradish peroxidase-conjugated goat anti-mouse IgG for 30 min. β-Actin was the control for sample loading and integrity. The antibody-reactive bands were revealed using an enhanced chemiluminescence kit (Amersham, Pittsburgh, PA, USA), and the bands were exposed to a Kodak radiographic film. The amount of polypeptide was quantitated by integrated densitometric analysis of the film (Kodak Gel Logic‐100 Imaging System).
6
Genotyping of NCF1 Gene Family
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The PCR co-amplification products of NCF1, NCF1B, and NCF1C were digested with BsrG1 and Pst1 (New England Biolabs, Frankfurt am Main, Germany) (37°C, 180 min), followed by enzyme inactivation (80°C, 20 min) (Figures 1 A–1C). The digestion fragments were developed in a 7.5% polyacrylamide (ratio 29:1) gel or a 5% (w/v) agarose gel stained with GelRed Nucleic Acid Gel Stain (Biotum, Fremont, CA, USA) and visualized using Gel Logic 100 Imaging System (Kodak, Eysins, Switzerland). Band intensities were quantified with ImageJ software.10 (link) For determination of the GTGT content (Figure 1 G), RFLP band intensities of 169-, 181-, and 201-bp BsrG1/Pst1 digestion products (Figures 1 D–1F, 2A, and 2 B) were size normalized by dividing band intensities by their length (number of base pairs). The size-normalized band intensity of the 169-bp band was divided by the sum of normalized band intensities of the 181- and 201-bp bands (Figure 1 G).
7
Polyplex Formation Characterization of siRNA-POD Complexes
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To assess the formation of polyplexes between the siRNA and the PODs,89 (link) Chol-POD, Palm-POD, PEG-PLGA-POD, and POD (Table 2 ) were mixed with siRNA in a PBS solution to give a final concentration of 35 μM for the PODs and 1 μM for the siRNA in a final volume of 10 μL and incubated for 30 min at room temperature. These formulations were then analyzed by electrophoresis on a 1% agarose, 0.5 × TBE (Tris-Borate-EDTA; UltraPure Agarose, Thermo Fisher, UK) gel for 40 min at 100 V and the gel visualized using the Gel Logic 100 Imaging System (Kodak). To determine the optimal ratio for formation of polyplexes, the same procedure was repeated for the QN-Palm-POD at four different molar ratios (35:1, 70:1, 140:1, and 200:1 POD:siRNA).
8
Molecular Identification of Culex pipiens Complex
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Morphological identification showed that the Cx. pipiens complex was the most common of all Culex species in the study area. Further molecular identification was conducted using PCR amplification to differentiate two members of the Cx. pipiens complex (i.e. Cx. pipiens pipiens and Cx. quinquefasciatus). This PCR targets the acetyl-cholinesterase-2 locus (ace-2). The ace-2 locus was amplified using primers B126, ACEquin, and ACEpip as previously described by Smith & Fonseca [47 (link)].
DNA was extracted from 280 specimens, randomly selected from the morphologically identified Cx. pipiens complex. A total of 5 µl of extracted genomic DNA per sample was amplified in a 20 µl reaction mix containing 1× PCR buffer, 250 µM dNTP, 2 mM MgCl2, 0.4 µM of universal primer and ACEquin, 0.2 µM of ACEpip, and 1 unit of Taq DNA polymerase overlaid by a drop of mineral oil. After PCR amplification, 10 µl of the DNA fragments were separated by electrophoresis on a 2.5% agarose gel stained with 0.5 µg/ml ethidium bromide and compared against a 100-bp DNA marker included in the gel. Separated DNA fragments were photographed under ultraviolet light using Kodak Gel Logic 100 imaging system and scored as Cx. pipiens pipiens (610 bp) or Cx. quinquefasciatus (274 bp).
DNA was extracted from 280 specimens, randomly selected from the morphologically identified Cx. pipiens complex. A total of 5 µl of extracted genomic DNA per sample was amplified in a 20 µl reaction mix containing 1× PCR buffer, 250 µM dNTP, 2 mM MgCl2, 0.4 µM of universal primer and ACEquin, 0.2 µM of ACEpip, and 1 unit of Taq DNA polymerase overlaid by a drop of mineral oil. After PCR amplification, 10 µl of the DNA fragments were separated by electrophoresis on a 2.5% agarose gel stained with 0.5 µg/ml ethidium bromide and compared against a 100-bp DNA marker included in the gel. Separated DNA fragments were photographed under ultraviolet light using Kodak Gel Logic 100 imaging system and scored as Cx. pipiens pipiens (610 bp) or Cx. quinquefasciatus (274 bp).
9
Polymer-Coated siRNA Nanoparticles for Delivery
10
Denaturing Protein Electrophoresis Protocol
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Electrophoresis was performed on denaturing and reducing polyacrylamide gels at 12% (30%T,2.67%C) [31 (link)]. Fifteen µL of the sample containing the precipitated proteins (1 µg/µL) and 5 µL of Laemmli sample buffer 4× (Bio-Rad, cat no. 1610747) were heated to 95 °C for 5 min and placed in each well of the gel. Precision Plus ProteinTM All Blue molecular Weight marker (Bio-Rad, cat. no. 1610373) was used. The gels were run in an electrophoresis chamber (MiniPROTEAN III, Bio-Rad) at 90 volts for 180 min and 20 mA (Bio-Rad, PowerPac Basic). After electrophoresis, the gels were washed with sterile distilled water, stained with Coomassie brilliant blue (Bio-Rad cat. no. 1610400) for 1 h, and destained with a solution (20% methanol (J.T. Baker®, USA), 15% acetic acid (J.T. Baker®, USA) and 65% deionized water) for 12 h. Gels were analyzed in the Gel Logic 100 imaging system (Kodak, USA) to establish the electrophoretic profile of protein bands.
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