Multigene gradient

Manufactured by Labnet

Sourced in United States

The Multigene Gradient is a precision thermal cycler designed for accurate and efficient DNA amplification. It features a gradient function that allows for simultaneous optimization of annealing temperatures across multiple samples, enabling researchers to quickly determine the optimal conditions for their PCR experiments.

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Lab products found in correlation

Gotaq flexi, by Promega (2 mentions) Taq pcr pre mix, by Solgent (1 mentions) Higene genomic dna prep kit, by Biofact (1 mentions) Isolate 2 plant dna kit, by Meridian Bioscience (1 mentions) Enduro gds, by Labnet (1 mentions) Taq dna polymerase, by Promega (1 mentions) Dreamtaq green buffer, by Thermo Fisher Scientific (1 mentions) Dntp mix, by Jena Biosciences (1 mentions) Dreamtaq dna polymerase, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

MultiGene Gradient Thermal Cycler (7 citations) Labnet MultiGene 96-well Gradient Thermal Cycler (6 citations) MultiGene (5 citations) MultiGene Gradient System (2 citations)

6 protocols using multigene gradient

Microsatellite Genotyping of Jatropha Genus

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Eleven primers previously described to be polymorphic for the jatropha genus were selected for this study. A detailed description of primers with their motif and sequence is shown in Table II.
For primer amplifications, the reaction was performed with the final volume of 25 µL containing 50 ηg of DNA, 6 µL of 5X reaction buffer, 1.5 mM MgCl 2 , 200 μM of each dNTP, 0.2 µM of each primer (Sigma, USA) and 2U of Taq DNA polymerase (Go Taq Flexi, Promega, USA). Samples were amplified in a gradient thermal cycler (Gradient Multigene, Labnet International, USA) using a touchdown program, with initial denaturation at 94°C for 5 min, followed by 8 denaturation cycles at 94°C for 50s, with the annealing temperature decreased 1ºC in each cycle from 62 to 55°C for 45s and 72°C for 1 min. This step was followed by 33 amplification cycles with denaturation at 94°C for 50 s, annealing at 54°C for 45 s and extension at 72°C for 1 min. Finally an amplification was concluded with final extension at 72°C for 10 min.
The amplified PCR product was run on a 6% denaturing polyacrylamide gel using 60W power for variable duration according to the expected size of the alleles. Then the resulting gels were stained with silver nitrate according to the method described by Creste et al. (2001) . After drying overnight at room temperature, the gels were photographed on a UV light box.

Santos D.N., Ferreira J.L., Setotaw T.A., Cançado G.M., Pasqual M., Londe L.C., Saturnino H.M, & Vendrame W.A. (2016). Genetic structure from the oldest Jatropha germplasm bank of Brazil and contribution for the genetic improvement. Anais da Academia Brasileira de Ciencias, 88(4).

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Genetic Diversity Analysis of Myrciaria dubia

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Fourteen ISSR markers were selected, based on their polymorphisms, and used in this study to analyze the genetic diversity among the Myrciaria dubia populations (Table 1). The DNA amplifications were performed in a thermal cycler gradient (Gradient Multigene; Labnet International, USA). The final reaction volume was 25 µL, containing 20 mM Tris-HCl pH 8.3, 1.5 mM MgCl 2 , 100 mM KCl, 0.10% Triton X-100, 200 µM each deoxyribonucleotide triphosphate (dNTP), 0.8 µM primer, 1.5 U enzyme GoTaq Flexi DNA polymerase (Prodimol, USA), and 50 ng DNA. In the polymerase chain reactions (PCR), the samples were initially subjected to 95°C for 2 min, followed by 40 amplification cycles consisting of 95°C for 45 s, 1 min annealing temperature (ranging from 47-52°C, depending on the primer used), and an extension step at 72°C for 2 min. After the cycles, the samples were kept at 72°C for 5 min for final extension. The PCR products were separated on a 1.5% agarose gel containing 0.2 mg/ mL EtBr (ethidium bromide) using horizontal gel electrophoresis run at 100 mV for at least 3 h in 1X TBE buffer. All gels were visualized and recorded using a UV image digitalizer (Uvitec, USA).

Nunes C.F., Setotaw T.A., Pasqual M., Chagas E.A., Santos E.G., Santos D.N., Lima C.G, & Cançado G.M. (2017). Myrciaria dubia, an Amazonian fruit: population structure and its implications for germplasm conservation and genetic improvement. Genetics and molecular research : GMR, 16(1).

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Molecular Identification of Bacterial Isolate AY001

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Total genomic DNA of AY001 was extracted using HiGene Genomic DNA Prep Kit (BIOFACT, Daejeon, Korea) according to the manufacturer’s instructions. Molecular identification of AY001 was performed by sequencing the amplified 16S rRNA region using primers 27F (5′-AGTTTGATCCTGGCTCAG-3′) and 1492R (5′-GTTACCTTGTTACGACTT-3′). PCR amplification was performed in a 30 μL reaction mixture containing 15 μL of 2X Taq PCR Pre-Mix (Solgent, Daejeon, Korea), 1 μL of primers mix, and 1 μL of gDNA template. PCR amplification was carried out using a thermal cycler (Multigene Gradient, Labnet, Edison, NJ, USA) by following amplification conditions: initial denaturation at 95 °C for 5 min, 33 cycles of 94 °C for 30 s, 58 °C for 30 s, 72 °C for 40 s, and final extension at 72 °C for 10 min. PCR product was purified, sequenced (Solgent, Daejeon, Korea), and analyzed with NCBI’s GenBank sequence database (http://www.ncbi.nlm.nih.gov, accessed on 1 March 2020) to identify the closest species relatives. The phylogenetic tree was constructed by MEGA-X software using the Maximum Likelihood method [21 (link)].

Heo A.Y., Koo Y.M, & Choi H.W. (2022). Biological Control Activity of Plant Growth Promoting Rhizobacteria Burkholderia contaminans AY001 against Tomato Fusarium Wilt and Bacterial Speck Diseases. Biology, 11(4), 619.

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ISSR-based Genetic Diversity Analysis

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DNA was extracted from young leaves using the Isolate II plant DNA kit (Bioline) following the manufacturer's instructions. DNA quality was examined by electrophoresis on 1 % agarose gels and DNA quantity was estimated using the spectrophotometer method. DNA 14 ISSR primers previously displayed reliable and polymorphic banding patterns were used in this study (Liu et al., 2007 , Athanasiadis et al., 2013) . The PCR amplifications were performed in a volume of 12.5µl, containing 12 ng of template DNA, 2.5 mM MgCl2, 0.8 mM dNTPs, 0.8 µM of each primer, 1X buffer and 0.75U of Taq DNA polymerase (promega, Madison, WI. USA). All PCR reactions were conducted in a DNA thermocycler (Multigene gradient, Labnet, NJ. USA) through 30 cycles, each consisting of 94°C for denaturation step (45 s), 45 s at the corresponding annealing temperature (°C), and a 72°C extension step (2 min), using the fastest available transitions between each temperature. Gradient PCR was used to adjust the annealing temperature of each primer. The last cycle was followed by a final extension for 7 min at 72°C. PCR products were separated by electrophoresis on 1.7% agarose with Ethidium Bromide in a TAE buffer and visualized by means of a Gel Doc system (Enduro TM GDS, Labnet). The sizes of amplification products were estimated using DNA marker (1 Kb, Invitrogen).

Bella Y.A., Bouda S., Harry M., & Haddioui A. (2021). Genetic variability of cultivated plum (Prunus domestica L. & Prunus salicina Lindl.) in Morocco assessed by ISSR markers. Australian Journal of Crop Science.

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PCR Amplification of DNA Targets

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Forward primer 5'- These primers were used in our last studies and by Miladi et al. [13, (link)39] (link).
Polymerase chain reaction (PCR) was performed in a reaction volume of 25 μl containing 2.5 µl 10X DreamTaq Green Buffer (Thermo Scientific), 100 µM dNTP Mix (JenaBioscience), 0.2 µM of both forward and reverse primers, 1 unit of DreamTaq DNA Polymerase (Thermo Scientific), and approximately 50 ng of DNA template. PCR reactions were conducted in a MultiGene gradient thermal cycler (Labnet International) using the following program: 95°C for 5 minutes, 35 cycles of 95°C for 30 seconds, 57°C (annealing) for 30 seconds, 72°C for 30 seconds, and finally 72°C for 10 minutes. PCR products were analyzed by electrophoresis on 1.5% agarose gel ethidium bromide stained.

Elabed H. (2021). Morphological Alterations and Virulence Gene Expression of Listeria Monocytogenes Cells in Response to Gamma Irradiation Treatments. Nutrition and Food Processing, 04(5), 01-09.

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Microsatellite Genotyping of Coffea arabica

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Forty-four microsatellite markers believed to be polymorphic for C. arabica species were tested in this study (Table S1).
PCR was performed in a volume of 25 µL, containing 50 ng DNA, 6 µL 5X reaction buffer, 1 mM MgCl 2 , 150 µM each dNTP, 0.1 µM primers, and 0.6 U Taq DNA polymerase (Go Taq Flexi, Promega, Madison, WI, USA). Reactions were carried out in a gradient thermocycler (Multigene Gradient, Labnet International, USA) using touchdown PCR with the following program: 94°C for a 2-min initial denaturing, followed by 13 cycles at 94°C for 30 s; annealing temperature 67°-55°C for 30 s, reducing 1°C in each cycle, extension at 72°C for 30 s. This was followed by 30 cycles of denaturation at 94°C for 30 s, 55°C for 30 s, and extension at 72°C for 30 s. A final extension occurred at 72°C for 8 min.
The amplification products were subjected to denatured gel electrophoresis on 6% polyacrylamide gel at 60 W power for variable times according to the size of the alleles. Next, bands were visualized using silver nitrate according to the method described by Setotaw et al. (2010) . The gel was then dried overnight at room temperature and photographed.

Pereira T.B., Setotaw T.A., Santos D.N., Mendes A.N., Salgado S.M., Carvalho G.R, & Rezende R.M. (2016). Identification of microsatellite markers in coffee associated with resistance to Meloidogyne exigua. Genetics and molecular research : GMR, 15(3).

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