Manipulating genome

• Genome is when all genes possessed by an individual organism ,exons are cod proteins made 2 percent of dna and introns are non coding dna Which is removed from mRNA before being transmitted
• satellite dna is a short sequence of dna which is repeated many time and 2 types of satellite dna are: minisatellite,micro satellite
• minisatellite,20-50 base pair repeated 50-100 times at more 100 location also known VNTR

  microsatellite,2-4 base pair repeated 5-15 times and known as STR and appear same location of chromosome
• The 5 stages of dna profiling -extracting dna and the digesting sample,separating dna fragments ,hybridisation,seeing the evidence
• extracting dna -dna is extracted from sample
• digesting sample-dna strands are cute in fragements using restriction endonuclease and they cut the dna at restriction sites and it makes 2 cuts and once though each strand of double helix and
• separate dna fragments -cut the dna fragments separated to form clear patterns using gel electrophoresis
• electrophoresis-dna fragments put in wells in agarose gel strips contains buffer solution to maintain constant ph and electric current is passed electrophoresis plate the fragments at cathode end move to anode need due to negative charge of phosphate group.small fragments move faster and then gel is placed in alkaline buffer to denature dna fragemnts and then souther blotting occur -occur in separating dna fragments
• southern blotting -place a nylon membrane all over gel and then all dna fragments slacked upon nylon sheets as its easier to process
• hybridisation-radioactive/fragement dye that is added to fragments on membrane and then dna probe sequence complementary to dna sequence and bind to the complementary strands of dna under particular condition
• seeing the evidence -if radioactive-x-ray are taken of paper/ membrane and if fluroscent the paper /memebrane under uv light
• the uses of dna profiling-used in fEileen of forensic science and performed on dna traces.used to prove paternity of child and immigration case and family relationships and it identify species to which organism belong and it can identify individuals what at risk to disease
• Polymerase chain reaction

  -sample dna is mixed with dna nucleotide,primer,mg,taq dna polymerase

  -mixture heated 94-96 degree breal h bonds between complementary nucleotide and denature double strand

  -mixture cooled 68 degree so primer anneal to one end of each dna strand

  taq dna polymerase can bind to end when double stand dna is

  -temp raised 72 degree keeping dna single strand

  -taq dna polymerase catalyse addition of dna nucleotide to single strand dna in 5-3 direction

  -new double strand is generated

  -process repeated
• polymerase chain reaction -occur when we extract dna its a natural process of dna replicated its amplified
• DNA taq polymerase-stands within higher temp used in pcr -attach nucleotide to synthesize dna

  restriction sites-sequence of 6-8 base pairs of dna that bind to restriction enzymes

  restrciation endonuclease-recognise specific dna sequence