Gel documentation system

Manufactured by Vilber

Sourced in France, Germany

The Gel documentation system is a laboratory equipment used for capturing and analyzing images of electrophoresis gels, such as those used in DNA, RNA, or protein analysis. The system comprises a camera, illumination source, and software for image acquisition and processing.

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Lab products found in correlation

Chemiluminescence reagent, by Thermo Fisher Scientific (2 mentions) Oligo dt 12 18 primer, by Thermo Fisher Scientific (1 mentions) E coli rnase h, by Thermo Fisher Scientific (1 mentions) Ptc 200, by Bio-Rad (1 mentions) Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Anti enos, by Cell Signaling Technology (1 mentions) Anti actin, by Cell Signaling Technology (1 mentions) Anti ace2, by R&D Systems (1 mentions) Hrp coupled secondary antibody, by Cell Signaling Technology (1 mentions) Qscript cdna synthesis kit, by Quanta Biosciences (1 mentions) Rotor gene q cycler, by Qiagen (1 mentions) Ecl select western blotting detection reagent, by GE Healthcare (1 mentions) Perfecta sybr green fastmix, by Quanta Biosciences (1 mentions) Rneasy plus kit, by Qiagen (1 mentions) Gelred fluorescent dye, by Biotium (1 mentions) Gotaq g2 green master mix, by Promega (1 mentions) Pierce biotin 3 end dna labeling kit, by Thermo Fisher Scientific (1 mentions) Type 1 agarose, by Merck Group (1 mentions) Stains all, by MP Biomedicals (1 mentions)

23 protocols using gel documentation system

Quantifying Collagen I Gene Expression

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Because we inserted human fibroblasts and bFGF into the nude mice, the gene expression of collagen type I in the implanted materials was examined using RT-PCR with human-specific primers. The results were normalized to the mRNA level of human GAPDH.
Total RNA was extracted by adding 0.5 ml of TRIzol® reagent (Invitrogen, Life Technologies, USA) to N₂-frozen nude mice tissues. Each μg of RNA was subjected to cDNA synthesis by using SuperScript™ Reverse Transcriptase II (Invitrogen) and oligo (dT)12–18 primers (Invitrogen) in a 20 μl reaction volume according to the manufacturer’s instructions, with the additional step of removing the RNA complementary to the cDNA using E. coli RNase H (Invitrogen). One microliter of each cDNA was then subjected to polymerase chain reaction (PCR) according to the following amplification profile: predenaturation at 94 °C for 40 s, amplification (denaturation at 94 °C for 40 s; annealing at 60 °C for 40 s; extension at 72 °C for 1 min) for 30 cycles, and a final extension at 72 °C for 10 min in a DNA thermal cycler (model PTC-200, MJ Research, Inc., MA, USA). For each of the PCR products, 10 μl was electrophoresed on a 1.5% agarose gel in the presence of ethidium bromide and visualized by the Gel Documentation System (Vilber Lourmat, France).

Lee S.Y., Park Y, & Hwang S.J. (2019). Effect of bFGF and fibroblasts combined with hyaluronic acid-based hydrogels on soft tissue augmentation: an experimental study in rats. Maxillofacial Plastic and Reconstructive Surgery, 41(1), 47.

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Gene Expression Analysis by qRT-PCR and Western Blotting

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Analysis of changes in gene expression was performed by quantitative real time PCR (qRT‐PCR). RNA was isolated using the RNeasy plus kit (Qiagen). Five hundred nanograms of mRNA were reverse transcribed using qScript cDNA synthesis kit (Quanta Biosciences). The resulting cDNA was analyzed on a RotorGene Q cycler (Qiagen) using PerfeCTa SYBR Green FastMix (Quanta Biosciences). For primer sequences see Table 1. The cycling program was 30 s 95°C for denaturing, followed by 45 cycles of 5 s 95°C, 15 s 56°C, and 10 s 72°C. Data analysis was performed by normalizing values to ß‐actin as a house‐keeping gene and comparing expression to normoxia (21% O₂) exposure using the ΔΔCt method. Values generated by qPCR are given as “fold change compared to 21% O₂.”
Protein levels were analyzed by SDS‐PAGE and western blotting. Transferred proteins were detected by primary anti‐ACE/CD143 (R&D Systems cat.nr. AF1513), anti‐ACE2 (R&D Systems cat.nr.AF3437), anti‐eNOS (Cell Signaling Technology cat.nr. 9,586), anti‐ß‐Actin (Cell Signaling Technology cat.nr. 3,700) and according to species‐specific HRP‐coupled secondary antibodies (Rockland, USA). Bands were visualized by chemiluminescence (ECL Select Western blotting Detection Reagent; Amersham; GE Healthcare) using a Gel Documentation System (Vilber Lourmat).

Wohlrab P., Johann Danhofer M., Schaubmayr W., Tiboldi A., Krenn K., Markstaller K., Ullrich R., Ulrich Klein K, & Tretter V. (2021). Oxygen conditions oscillating between hypoxia and hyperoxia induce different effects in the pulmonary endothelium compared to constant oxygen conditions. Physiological Reports, 9(3), e14590.

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Bacterial Identification by RAPD-PCR Analysis

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RAPD-PCR was performed for all the bacterial isolates to confirm their identity up to the species level. Four types of 10-mer random primers were used for RAPD analysis. The primers were named as primer A (5′-GTGATCGCAG-3′), primer B (5′-CTTTCGCTCC-3′), primer C (5′-CGCAGACCTC-3′), and primer D (5′-GAACTGGAGT-3′). PCR was conducted in a reaction mixture of 25 μL, containing DNA template (1.0 μL, 50 ng), 12.5 μL GoTaq^® G2 Green Master Mix (pH 8.3, 1.5 mM MgCl₂, 200 μM of each dNTP, DNA polymerase) (Promega; Madison, WI, USA), primer (2.5 μL) (Bionics, Seoul, Republic of Korea), and nuclease-free distilled water (9.0 μL). PCR was carried out at initial denaturation for 5 min at 94 °C, followed by 40 cycles [95 °C for 15 s (denaturation), 36 °C for 15 s (primer A and primer B), 38.4 °C for 15 s (primer C), or 28.1 °C for 15 s (primer D) (annealing), and 72 °C for 2 min (extension)], and a final extension at 72 °C for 4 min. The amplified RAPD-PCR product (10.0 μL) was resolved on a 1.2% agarose gel at 50 V, followed by staining with a GelRed^® fluorescent dye (Biotium, Fremont, CA, USA) for 10 min, and visualized using a gel documentation system (Vilber Lourmat, Marne-la-vallee, France).

Bahuguna A., Joe A.R., Kumar V., Lee J.S., Kim S.Y., Moon J.Y., Cho S.K., Cho H, & Kim M. (2020). Study on the Identification Methods for Effective Microorganisms in Commercially Available Organic Agriculture Materials. Microorganisms, 8(10), 1568.

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Quantifying AP-1 Binding Activity

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The binding activity of activator protein (AP)-1 in nuclear fractions was assayed using an electrophoretic mobility shift assay. AP-1 consensus double stranded oligonucleotide (5′-CGC TTG ATG ACT CAG CCG GAA-3′) (Santa Cruz Biotechnology, USA) was end-labeled with Pierce Biotin 3′ End DNA Labeling Kit (Thermo Fisher Scientific, USA). Briefly, 10 μg of nuclear protein samples were incubated with 100 fmol of oligonucleotides at room temperature for 20 min and separated on a 6% polyacrylamide gel. The specificity of binding was determined by 200-fold unlabeled oligonucleotide competition. The AP-1-DNA complexes were transferred onto 0.2 μm polyvinylidene fluoride membranes and incubated with chemiluminescence reagent (Thermo Scientific, USA) for 1 min and imaged in a gel documentation system (Vilber Lourmat, Germany). Band intensities were quantified by ImageJ Image Processing Software (NIH, USA).

Lee Y.Z., Yap H.M., Shaari K., Tham C.L., Sulaiman M.R, & Israf D.A. (2017). Blockade of Eosinophil-Induced Bronchial Epithelial-Mesenchymal Transition with a Geranyl Acetophenone in a Coculture Model. Frontiers in Pharmacology, 8, 837.

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R-Plasmid Screening via Alkaline Lysis

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The selected bacterial isolates were screened for R-plasmid by alkaline lysis method which described in Feliciello and Chinali, 1993 [14 (link)]. A 30 µl plasmid sample was electrophoresed through 0.7% Agarose (Type 1,
Sigma) with ethidium bromide (0.6µg/ml) in TE buffer at 120 V for 3 hours. The bands were visualized in a gel documentation system (Vilber Lourmat, France).

Rani P., Singh N., Bhaskar M, & Tandan N. (2022). Data on antibiotic resistance among indoor microbiome at Meerut, India. Bioinformation, 18(3), 293-298.

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Molecular Genotyping of Lentil Genotypes

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DNA was extracted using modified CTAB method [25 ] and was quantified using a spectrophotometer. Thirty SSR markers which were polymorphic between two genotypes that a showed contrasting response to alkalinity stress, along with additional 38 arbitrary SSR markers were used for screening 285 genotypes. These markers were selected based on earlier lentil reports published [16 (link), 26 (link), 27 (link)].
PCR amplifications were performed in 10μl reaction volume, consisting of 1 X PCR buffer, 1.5 mM MgCl₂ and 0.5 μM primers each of forward and reverse, 1 mM dNTP, 0.5 U Taq DNA polymerase and 50 ng template DNA. PCR cycling conditions were as follows: Pre-denaturation at 94°C for 3 min followed by 40 cycles of denaturation at 94°C for 30 sec, annealing at 55°C for 30 sec, elongation at 72°C for 1 min with a final extension at 72°C for 10 min. PCR amplified products were separated on 3% ultra high resolution agarose gels and documented using Vilber Lourmat Gel Documentation System.

Singh D., Singh C.K., Singh Y.P., Singh V., Singh R., Tomar R.S., Sanwal S.K., Karwa S., Mishra V.K., Sarkar S.K., Pal M., Kumar A., Yadav R.K, & Sharma P.C. (2018). Evaluation of cultivated and wild genotypes of Lens species under alkalinity stress and their molecular collocation using microsatellite markers. PLoS ONE, 13(8), e0199933.

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Oligonucleotide Stability in Cell Culture

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Oligonucleotide 1, oligonucleotide 3 or conjugate 3 in concentration 0.1 µg/µl were incubated in Dulbecco’s Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C for 48 h. Aliquots (10 µl) were taken at 0, 2, 4, 8, 24 and 48 h and reaction was quenched by adding 10 µl of 8 M urea and immediate freezing in liquid nitrogen. Samples were defrosted and analyzed in 12% PAAG/8 M urea using TBE as running buffer. Then electrophoresis gels were stained with Stains-All (MP Biomedicals, USA) and photographed using gel documentation system (Vilber Lourmat, France).

Patutina O.A., Bazhenov M.A., Miroshnichenko S.K., Mironova N.L., Pyshnyi D.V., Vlassov V.V, & Zenkova M.A. (2018). Peptide-oligonucleotide conjugates exhibiting pyrimidine-X cleavage specificity efficiently silence miRNA target acting synergistically with RNase H. Scientific Reports, 8, 14990.

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Isozyme Profiling of Maize Seedlings

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Four isozymes, leucine-aminopeptidase (LAP), esterase (EST), peroxidase (PER), and catalase (CAT), were used in this experiment. The dried leaves of ELF exposed and nonexposed maize seedling were separately milled and defatted according to methods described by [22 (link)]. Approximately 0.4 g powdered seed was crushed with acid washed sand and 400 mL extraction buffer. Extraction buffer consisted of 0.1 M Tris-HCl (pH 7.5) containing 20% sucrose as described by [27 ]. The samples were then centrifuged at 15000 g for 15 min at 4°C; supernatants were collected and used directly for isozyme analyses in a separate vial. Each sample was applied to vertical polyacrylamide gel electrophoresis (4.5% stacking, 9% separating gel) using a mini gel apparatus in Tris-glycine (pH 8.3) buffer as described by [27 ]. Visualization of enzyme activity after electrophoresis was achieved by histochemically staining the gels. The gels were stained for LAP, EST, PER, and (CAT) separately with specific activity stain solutions as described by [28 (link)–31 (link)], respectively. Gels were photographed using the Vilber Lourmat gel documentation system.

AL-Huqail A.A, & Abdelhaliem E. (2015). Evaluation of Genetic Variations in Maize Seedlings Exposed to Electric Field Based on Protein and DNA Markers. BioMed Research International, 2015, 874906.

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Polymerase Chain Reaction Protocol

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Polymerase chain reaction (PCR) amplifications were performed in a reaction volume of 10 μl containing 1 μl of genomic DNA (25 ng/μl) as template, 1.0 μl each of forward and reverse primers (10 ng/μl), 1 μl of dNTPs (10 mM), 0.5 units of Taq DNA polymerase, 1.0 μL of 10X PCR buffer and the rest was milliQ water. After initial denaturation at 94°C for 3 min, PCRs were run for 30 cycles with a denaturation step of 1 min at 94°C, annealing for 1 min at 55-60°C, and extension at 72°C for 2 min. A final extension was followed at 72°C for 10 min. The amplified PCR products (10 μl) were run on 1.5% (w/v) agarose gel at 120 V in 1 X TBE buffer. The size of the fragments was estimated using a 100 bp ladder (Genei, Bangalore) as size marker. The gels were photographed using the Gel Documentation System (VilberLourmat, France). All PCR reactions were done in triplicate to ensure the reproducibility and reliability of the results. Only highly reproducible and polymorphic primers were included in the study.

Mohanavel V., Yesudhas A.S., Sharma A., Ramasamy A., Muthu Arjuna Samy P., Subramanian M, & Muthusamy R. (2021). Haplotype and diversity analysis of indigenous rice for salinity tolerance in early-stage seedling using simple sequence repeat markers. Biotechnology Reports, 31, e00666.

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Genotyping IRS-2 G1057D Polymorphism

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Genomic DNA was extracted from 200 µl of peripheral blood by High Pure DNA isolation Kit (Qiagen, Inc., Chatsworth, CA) following then manufacturer's instructions. A polymerase chain reaction (PCR)-based restriction fragment-length polymorphism (RFLP) method was used for genotype IRS-2 G1057D polymorphism, which removes the Hae-II restriction enzyme site. The PCR was performed in a 25 µl volume containing 20 ng genomic DNA, 10×PCR buffer with 1.5 mM MgCl₂, 0.25 mM dNTPs, 10% dimethylsulphoxide, 0.5 units of Taq polymerase (Fermantas, MBI) and 5 pmol of each primer, IRS-2F (5’ GCT CCC CCA AGT CTC CTA A 3’) and IRS-2R (5’ CTC AGC CTC TTC ACG CCC 3’). The PCR thermal cycling conditions were an initial melting period at 95°C for 2 min; then 35 amplification cycles of 95°C for 45 s, 62°C for 45 s and 72°C for 45 s; and a 7-minute extension step at 72°C. The PCR products were checked on 1.5% agarose gel for the assay completion, and then the PCR products of 375 base pairs (bp) were digested with restriction enzyme Hae-II by overnight incubation at 37°C. The digestion products were electrophoresed on 3% agarose gel and visualised by staining with ethidium bromide and evaluated using the gel documentation system (Vilber-Lourmat, Cedex, France).

Yukseloglu E.H., Celik S.K., Kucuk M.U., Yalin E., Ozkal S.S., Ates C., Berkoz M., Yalin S, & Ates N.A. (2014). IRS-2 G1057D polymorphism in Turkish patients with colorectal cancer. Przegla̜d Gastroenterologiczny, 9(2), 88-92.

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