CONTENT : Defination of blotting History Southern blotting Northern blotting Western blotting Application DEFINATION : Transfer of DNA or RNA or protiens fractinated by gel electrophoresis to a nitrocellulase paper or nylon mesh for separation , identification and analysis is called blotting or blot transfer . It is carried out by allowing a liquid to seep through electrophoresed gel , nitrocellulose paper and a pile of filter papers. They are very important for hybridisation experiments to determine the DNA , mRNA or protiens in ...
What is blotting? Blots are techniques for transferring DNA , RNA and proteins onto a carrier so they can be separated, and often follows the use of a gel electrophoresis. The Southern blot is used for transferring DNA, the Northern blot for RNA and the western blot for PROTEIN. TYPES OF BLOTTING TECHNIQUES Blotting technique Southern Blot Northern Blot Western blot It is used to detect DNAI.t is used to detect RNAIt. is used to detect protein. SOUTHERN BLOTTING &bull ...
Blotting • All techniques use electrophoresis to separate. • All are based on hybridization technique • Difference in techniques lies in the target • Four applications –Western –Southern –Northern –Southwestern 08/27/2022 2 Hybridization The binding between ss labeled probe to a complementary nucleotide sequence on the target DNA. Degree of hybridization depends on method of probe labeling (radioacitve or non-radioactive system e.g. biotin-avidin. 08/27/2022 3 Probes • Labeled material to detect a target. • For ...
Southern Blot (1975) • Bu yontemde izole edilen DNA uygun bir restriksiyon enzimi ile kesilir ve belirli say ve uzunlukta DNA frangmentlerinin oluumu salanr. Agaroz jel elektroforez yontemi ile jel uzerinde buyukluklerine gore birbirinden ayrlan bu fragmentler once yuksek pH ile denature edilip blot yontemi ile nitroseluloz filtre uzerine gecirilir. Southern Blot (1975) • 32 p (ya da baka bir radyoaktif olmayan nukleotit analou) ile iaretli denature edilmi ve incelenecek bolgeye ozgu prob, filte uzerinde tutulan fragmentler ile uygun koullar ...
DNA Extraction Gel Electrophoresis • Because nucleic acids are negatively charged ions at neutral or basic pH in an aqueous environment, they can be mobilized by an electric field. Gel electrophoresis is a technique used to separate molecules on the basis of size, using this charge. PCR • PCR is used to amplify a specific sequence of DNA. Reverse Transcriptase PCR DNA fragments can also be amplified from an RNA template in a process called reverse transcriptase PCR ...
DNA EXTRACTION METHODS DNA extraction is used to isolate Types of DNA Mitochondrial DNA Genomic DNA Plasmid DNA How Can We Recover DNA From a Variety of Sources of Biological Evidence? Blood, Tissues, Saliva, Urine, Hair (w/Root & Shaft), Teeth, Bone, Semen, Cigarette Butts, Envelope & Stamps, Fingernail Clippings, Chewing Gum, Feces Purpose of DNA Extraction To obtain DNA in a relatively purified form which can be used for further investigations such as: PCR (polymerase chain reaction) RFLP ...
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