Principle of CAPS and dCAPS Markers

Maxam and Gilbert Sequencing Method or Chemical Cleavage Sequencing Method is a first generation sequencing method cleave specific DNA molecule by certain chemicals and subsequent cleavage at specific bases as (G), (A+G), (C) and (C+T) which correspond to radiolabeled fragments, Maxam and Gilbert Sequencing Method initiate by DNA extraction and denature double strand DNA molecule to single fragments and radiolabel single fragments on 5’ ends using gamma 32P ATP After that cleave single fragments at specific positions using four chemicals reactions to generate breaks in specific nucleotides as follow: Tub- 1 contain dimethyl sulfate to break guanine nucleotide and Tube-2 contain dimethyl sulfate and formic acid to break purine nucleotides and Tube-3 contain hydrazine to break pyrimidine nucleotide and Tube-4 contain hydrazine and sodium chloride to break cytosine nucleotide and incubate four reactions tubes with piperidine to break modified strands and subject reactions side by side in denature polyacrylamide gel electrophoresis to differentiate fragments size according to radiolabels and autoradiography gel on X-ray film to read gel bands from bottom to top, Bands present in cytosine and pyrimidine nucleotides refer to cytosine nucleotide and bands present in pyrimidine nucleotides refer to thymine nucleotide and bands present in guanine and purine nucleotides refer to guanine nucleotide and bands present in purine nucleotides refer to adenine nucleotide, Maxam and Gilbert Sequencing Method use purified DNA directly in sequencing process to identify DNA modification without require PCR and consider accurate and suitable in DNA foot printing studies, Maxam and Gilbert Sequencing Method consider labor and costly and time consuming method and require hazardous chemicals as radioactive labels and Read short sequence length (50 to 500) bp, Maxam and Gilbert Sequencing Method use in DNA Structure Studies and DNA Modification Studies and DNA Interaction Studies and DNA Footpriting Studies and DNA Fingerprinting Studies (Watch Related Video in #geneticteacher) #geneticteacher