To study polymorphism in the chickpea core collection, PCR amplification was carried out in a Universal Gradient thermal cycler with 96 wells using a 20-μL reaction mixture: 2 μL of template DNA (25 ng/μL), 10 × PCR buffer (1.8 mM, MgCl2, 10 mM Tris-HCl, 50 mM KCl), 2.5 mM dNTPs (Molecular Biology for Life Science, Fermentas, Lithuania, USA), 5 μM each of forward and reverse primer and 5U of Taq DNA polymerase (Sigma Aldrich USA. The thermal cycler (Peqlab) was programmed as follows: initial denaturation for 5 min at 94 °C followed by 40 cycles of denaturation for 30 s (s) at 94 °C, annealing at temperature specific for each primer pair for 30 s and extension at 72 °C for 30 s. The final extension was allowed for 10 min at 72 °C and storage at 4 °C until further use. The resulting PCR products were run on 10% Poly-acrylamide Gel Electrophoresis (PAGE) using a Dual Gel Vertical Electrophoresis System (Peqlab) and silver stained for manual visualization of bands on gel documentation system.
Dual gel vertical electrophoresis system
Manufactured by Avantor
The Dual Gel Vertical Electrophoresis System is a laboratory equipment used to separate biomolecules, such as proteins or nucleic acids, based on their size or charge through the process of electrophoresis. It provides a reliable and consistent method for conducting vertical gel electrophoresis experiments.
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Lab products found in correlation
2 protocols using dual gel vertical electrophoresis system
1
Analyzing Chickpea Genetic Diversity
2
Assessing Chickpea Genetic Diversity
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To study polymorphism in the chickpea core collection, PCR amplification was carried out in a Universal Gradient thermal cycler with 96 wells using a 20-μL reaction mixture: 2 μL of template DNA (25 ng/μL), 10x PCR buffer (1.8 mM, MgCl2, 10 mM Tris-HCl, 50 mM KCl), 2.5 mM dNTPs (Molecular Biology for Life Science, Fermentas, Lithuania, USA), 5μM each of forward and reverse primer and 5U of Taq DNA polymerase (Sigma Aldrich USA.
The thermal cycler (Peqlab) was programmed as follows: initial denaturation for 5 min at 94°C followed by 40 cycles of denaturation for 30 seconds (s) at 94 °C, annealing at temperature specific for each primer pair for 30 s and extension at 72 °C for 30 s. The final extension was allowed for 10 min at 72 °C and storage at 4 °C until further use. The resulting PCR products were run on 10% Poly-acrylamide Gel Electrophoresis (PAGE) using a Dual Gel Vertical
Electrophoresis System (Peqlab) and silver stained for manual visualization of bands on gel documentation system.
The thermal cycler (Peqlab) was programmed as follows: initial denaturation for 5 min at 94°C followed by 40 cycles of denaturation for 30 seconds (s) at 94 °C, annealing at temperature specific for each primer pair for 30 s and extension at 72 °C for 30 s. The final extension was allowed for 10 min at 72 °C and storage at 4 °C until further use. The resulting PCR products were run on 10% Poly-acrylamide Gel Electrophoresis (PAGE) using a Dual Gel Vertical
Electrophoresis System (Peqlab) and silver stained for manual visualization of bands on gel documentation system.
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