Explore the the green world with us
Gene cloning is a process in which multiple copies of genetic material, such as genes, are created. To do this, scientists utilize vectors – specific strands of DNA that can be inserted into cells and transport the desired genes with them. Through gene cloning, researchers have been able to gain valuable insight into genetics and … Read More “Vectors for Gene Cloning” »
Performing a restriction digest in the laboratory Digest a sample of λ DNA (concentration 125 μg ml−1) with HindIII DNA – The amount of DNA required for restriction digestion varies depending on the experiment and its desired results. – To digest 2 μg of λ DNA -16 μl of the DNA sample is needed Buffer … Read More “Manipulation of Purified DNA” »
Ti-plasmid are widely utilized to introduce DNA into plant cells. but natural form of Ti plasmids are not suitable for as cloning vector because The size of the Ti plasmids is 150kb-200kb, it is extremely difficult to manipulation. Binary vector Consist a helper plasmid and a donor plasmid. Helper plasmid (binary cloning vector)contains disarmed Ti … Read More “Plasmid Vectors for plant cells” »
Recombinant DNA Technology is the process that isolation and manipulation of DNA sequences. it involves cutting, combining of DNA from different sources and create new DNA combination/genes. Gene Cloning Gene cloning is the process of generation of genetically identical copy of cell or an organism. A fragment of DNA, that gene of interest on cloning … Read More “Recombinant DNA Technology” »
Blotting Techniques mainly have four main steps Electrophoretic separation of protein/nucleic acid fragments in the sample Transfer to and immobilization on a solid support Binding of probe to the target molecule on membrane Visualization of bound probe Southern blotting – Allows DNA fragments to be identified Northern blotting – Allows RNA fragments to be identified … Read More “Blotting Techniques” »
Electrophoresis − the migration of a charged particle under the of an electric field. Migration rate/separation depends on: size, shape, charge The charge per unit length in any given fragment of DNA is the same since mobility of DNA molecules during gel electrophoresis The small DNA fragments move more easily through the gel pores. Requirements … Read More “Electrophoresis of Nucleic Acids.” »
DNA and RNA quantitation is performed using various methods. Absorbance/spectrophotometric methods Gel electrophoresis – Agarose gel electrophoresis Fluorometric methods Absorbance method (UV spectrophotometer, NanoDrop) At 260 nm nucleic acids absorbs light most strongly At 280 nm proteins are absorbs light most strongly At 230 nm other chemical contamination absorbs light most strongly. DNA purity (protein … Read More “DNA/RNA QUANTIFICATION” »
Plasmid DNA may appear in several conformations. Covalently closed circular DNA (cccDNA) – both strands of the plasmids DNA are intact. (Supercoiled DNA, Relaxed circular DNA) Open circular DNA – One nucleotide stand is broken Super coiled DNA is move faster than open circular and genomic DNA because it compact conformation. Plasmid DNA extraction methods. … Read More “Plasmid DNA extraction.” »
RNases enzyme is one of the main problem of the RNA extraction. that degraded the RNA because RNases is very stable, found every everywhere, active enzyme that do not required any co-factors and maintain activity even during the autoclaving. Avoiding RNase Contamination RNase inhibitors are added to the extraction buffer to inhibit the endogenous RNase: … Read More “RNA Extraction.” »
The key steps of DNA extraction is give below. Sample disruption Clearing cell debris Purification Concentration DNA Sample disruption. rapid and complete disruption of cells in sample releasing DNA into lysate. there are several methods to disruption of the cell wall. Physical methods – grinding with mortar and pestle under liquid nitrogen. Chemical methods – … Read More “DNA extraction” »
