Southern blotting onto nylon or nitrocellulose membrane ( Brown, 2001) Then, the membrane is placed in a solution of labeled RNA, single-stranded DNA, or oligodeoxynucleotide complementary to the blot transferred DNA bands to be detected. After transfer, the DNA is fixed onto the membrane. Nitrocellulose or nylon serves as the membrane material. The DNA molecules are run on the membrane. The hybridization membrane is sandwiched between the gel and paper towels which draw the transfer buffer through the gel by capillary action. The agarose gel for southern blotting is mounted on a filter paper dipped in a reservoir containing transfer buffer. The desired DNA is detected using a labeled probe complementary to the desired DNA. The nitrocellulose filter is sandwiched between the gel and the stack of paper towels which draws the transfer buffer from the gel through capillary action. The technique involves the transfer of electrophoresed DNA from the electrophoresis gel to a nitrocellulose membrane. The DNA fragments are separated based on their size and charge during electrophoresis. Southern blotting is based on the principle of separation of DNA fragments by gel electrophoresis followed by the identification by labeled probe hybridization. In addition to its use for DNA analysis, the immunologists have long used the southern blotting for gene identification in somatic rearrangements and transgene studies. Furthermore, southern blotting is used to identify methylated sites in particular genes. Southern blotting could be used for homology-based cloning by exploring the amino acid sequence of the protein product of the gene of interest. Sequences hybridizing with the hybridization probe are further analyzed to obtain the full-length sequence of the gene of interest. These oligonucleotides are chemically synthesized, radiolabeled, and then used for the cloned DNA fragments. Oligonucleotides, similar to the target sequence are designed. After immobilization, the DNA fragments could be subjected to hybridization analysis, enabling the bands with sequence similarity to a labeled probe. The method involves the transfer of DNA fragments from an electrophoresis gel to a membrane causing immobilization of the DNA fragments and bands are produced. Southern, who developed the technique in the 1970s. Southern blotting is a hybridization technique used for deoxyribose nucleic acid (DNA). Balances, Scales and Weighing Equipment.
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