![]() ![]() To visualize the DNA bands present in the gel, stains such as methylene blue and ethidium bromide are used. The buffer is a mixture of organic and inorganic salts that helps to conduct the electric current between the positive and negative terminals. The solid matrix controls the rate of migration of the molecules based on the size of the particles and the concentration of the gel. Here the agarose gel is the solid matrix. The DNA are negatively charged particles that are attracted towards the opposite charge under the influence of electric field. Gel electrophoresis is the most powerful technique for separating the biomolecules. These DNA fragments thus obtained are separated using the Agarose gel electrophoresis. The number of DNA fragments and the size of the DNA fragments depend upon the action of the restriction enzyme. The restriction mapping is used to identify the plasmids. This restriction digestion is used for two purposes: Restriction mapping and specific DNA cleavage for the production of new constructs. Each restriction enzyme has a unique code and it cuts the DNA into fragments with either sticky or blunt ends.Ī restriction map gives us the location where the restriction enzyme cuts the DNA. The pieces of DNA that remain after the digestion with the restriction enzymes are called as restriction fragments. The recognition sites for these restriction enzymes are as follows: EcoRI recognition site = G|AATTC Bam H1 recognition site = G|GATCC C TTAA |G C C T A GIG and Pst 1 recognition site = CTGCAIG GIACGTC (Siwach and Singh 2007). The most common restriction endonucleases are EcoR1, BamH 1 and Pst1.Īll these restriction enzymes have sticky ends. There are about 200 different restriction enzymes. Restriction endonucleases are the enzymes that cut the DNA at the specific sequences. These vectors are of two types: expression vectors (expression of the cloned gene to give the desired protein) and cloning vectors (produce millions of copies of cloned DNA). These vectors replicate inside the host cell along with the inserted DNA. This DNA is now called as recombinant DNA. ![]()
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