RNA Polymerase: Synthesis RNA from DNA Template
RNA polymerase is an enzyme that catalyzes the synthesis of RNA from ribonucleoside triphosphates in the presence of a DNA template and divalent cation, such as Mg2+ or Mn2+. The RNA molecules are synthesized complementary and antiparalel to one of the DNA strands in a 5, to 3, direction. The ribonucleotides are covalently joined together by internucleoside 3, to 5, phosphodiester bonds with concomitant release of inorganic pyrophosphate. Continue reading ‘RNA Polymerase: Synthesis RNA from DNA Template’
RNA Polymerase: Synthesis RNA from DNA Template
RNA polymerase is an enzyme that catalyzes the synthesis of RNA from ribonucleoside triphosphates in the presence of a DNA template and divalent cation, such as Mg2+ or Mn2+. The RNA molecules are synthesized complementary and antiparalel to one of the DNA strands in a 5, to 3, direction. The ribonucleotides are covalently joined together by internucleoside 3, to 5, phosphodiester bonds with concomitant release of inorganic pyrophosphate. Continue reading ‘RNA Polymerase: Synthesis RNA from DNA Template’
Restriction Enzyme Storage
Restriction enzymes are endonucleases that recognize specific double stranded DNA sequences and cleaves the DNA in both strands. Restriction enzymes used in cloning are site specific. Cloning involves cleaving both the vector and genomic DNAs with a restriction endonuclease which yields compatible sticky ends, and then using those cohesive ends to recombine the DNAs into a construct or recombinant molecule. The vector is cleaved in one location, while the genomic DNA is extensively digested into a pool of fragments. The quality of restriction enzyme determines the successful of cloning process. Therefore, its storage is very important to be known by us.
Knowing More About Nitrification
Nitrification is the biological formation of nitrate from sequentially oxidizing ammonium with the intermediate formation of nitrite. Those two oxidative reactions are catalyzed by ammonium oxidizers and nitrite oxidizers.
What is Isoelectric Point?
The isoelectric point (pI) is the pH at which any given protein has an equal number of positive and negative charges, in other word the protein has no charge or neutral. At a pH below the isoelectric point, proteins carry a net positive charge, and above the isoelectric point protein has a net negative charge*
RNA Polymerase: Synthesis RNA from DNA Template
RNA polymerase is an enzyme that catalyzes the synthesis of RNA from ribonucleoside triphosphates in the presence of a DNA template and divalent cation, such as Mg2+ or Mn2+. The RNA molecules are synthesized complementary and antiparalel to one of the DNA strands in a 5, to 3, direction. The ribonucleotides are covalently joined together by internucleoside 3, to 5, phosphodiester bonds with concomitant release of inorganic pyrophosphate. Continue reading ‘RNA Polymerase: Synthesis RNA from DNA Template’
Restriction Enzyme Storage
Restriction enzymes are endonucleases that recognize specific double stranded DNA sequences and cleaves the DNA in both strands. Restriction enzymes used in cloning are site specific. Cloning involves cleaving both the vector and genomic DNAs with a restriction endonuclease which yields compatible sticky ends, and then using those cohesive ends to recombine the DNAs into a construct or recombinant molecule. The vector is cleaved in one location, while the genomic DNA is extensively digested into a pool of fragments. The quality of restriction enzyme determines the successful of cloning process. Therefore, its storage is very important to be known by us. Continue reading ‘Restriction Enzyme Storage’
Knowing More About Nitrification
Nitrification is the biological formation of nitrate from sequentially oxidizing ammonium with the intermediate formation of nitrite. Those two oxidative reactions are catalyzed by ammonium oxidizers and nitrite oxidizers.
The stoichiometric equation for the oxidation of ammonium to nitrite by ammonium oxidizers is as follows : Continue reading ‘Knowing More About Nitrification’
What is Isoelectric Point?
The isoelectric point (pI) is the pH at which any given protein has an equal number of positive and negative charges, in other word the protein has no charge or neutral. At a pH below the isoelectric point, proteins carry a net positive charge, and above the isoelectric point protein has a net negative charge. Continue reading ‘What is Isoelectric Point?’
Separation of DNA Fragments Using PAGE Method
This method is able to separate DNA fragments with the size of as small as 10 bp and up to 1 kb with the resolution of as little as 1 bp. While agarose gel electrophoresis is only able to separate DNA fragments with the bigger size that PAGE does or in the size range of 100 nucleotides to around 10 – 15 kb. Continue reading ‘Separation of DNA Fragments Using PAGE Method’
Quantitation of Biosurfactant
Biosurfactants can be quantified by surface and interfacial tension. This is a generic quantitation and thus does not distinguish among different types of surfactants that may be present. Biosurfactants can be compared in terms of the amount they reduce surface or interfacial tension, and the critical micelle concentration (cmc), which is the lowest surfactant concentration above which no further decrease in surface tension or interfacial tension takes place. Continue reading ‘Quantitation of Biosurfactant’
Purification of Plasmid DNA
After the initial characterization, it is possible to purify further some or all of the plasmid DNAs by RNase digestion and extraction with organic solvents. This further purified DNA is suitable for techniques such as DNA sequencing, subcloning or the production of gene probes. In order to purify plasmid DNA after the isolation process, any residual RNA and contaminating protein are removed. This purification step involves two main steps, which are, first, removing residual RNA by using RNase in order to digest RNA and, second, extract contaminating protein using organic solvents, phenol-chloroform. Continue reading ‘Purification of Plasmid DNA’