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> - DNA structure > - RNA struc...

> - DNA structure > - RNA structure A-T-G- c (Hydrogen Bonding A-U + G- e (Hydrogen Bonding · · Antiparrallel Right-Handed Helix ↑ · + -- -- Ho it No H > - Prokaryotic Gene Structure > - Eukaryotic gene structure no introns or exons just genes Introns and Exons · · ; · operons Single · bidirectional promoter (sometimes) El E I - Genez #Genez/ Genesi Genet ENenea/Genesi - anney operon operon Techniques for studying DNA > - DNA Digestion ↑ Genomes = ↓ smaller fragments = RE that have smaller recognition site 1) Restriction Enzymes (RE) - Restriction Endonucleases cut dsDNA at specific Sites recognition · make either blunt or sticky ends · · Type #P(Palindromic) : Most common RE (4-8 bp recog) - Iso chizomers but respond · REswl same recognition sites differently to methylation state Ex DAM (cuts when sequence is unmethylated) DpnI (cuts when seg is methylated + · : > - separation DNA 1) Agarose Ge Electrophoresis * Problem : DNA Smear : Too many fragments = No clear Separates aspNA fragments by - size separation 2) Pulsed field Electrophoresis - Separates large dsDNA molecules > - DNA Amplification > - Verification Techniques I PCR (Polymerase chain repeat 1) Agarose get : Check to see if Agarose get S steps Denaturation (940) : weigh corresponds PCR product size - Annealing (60%) Repeat 30 times Elongation (720c) 2) Restriction Analysis : RE site in product will be targeted if target is correct materials Template dsDNA - : Primer pairs 3) Nested PCR : Primers that bp inner DNA are Heat resistant DNA polymerase made (nest PCR product = PCR product) ANTPs 1) Sequence Per product  2) RT PCR (MRNA-CONE) - steps : Synthesize CDNA strand PCR procedure - materials : Template SSCDNA poly T primer Heat resistant RT dNTDs sample) Intelling curve 3) qPCR (measure relative+ absolute MRNA in I Pur product 1 peak (Expected) - N fluorescence = mRNA to start = Quicker plateau most mRNA E DNA Analysis > - D) (DNA) Southernblot -most stable, other options (CDNA , RNA , oligomer) -Agarose get transfered to blot Tagged w T probe for visualization - Transfer via capillary action (weight or Electroblotting (current) RNA 2) Northernblot (RNA) 3) Western blot (proteins) Protein -Y wl antibody Tagged ↓! - 4) Southernwestern blot 2) Northernwestern blot DNA - Electrophoretic mobility shit Assay : Study interactions blw DNA (probe) /moteins DNA library properation al Fragmentation (Sonrication/R E GONE Collect DNA from organism or RTMRWA Into CANA /more than 1 copy of. 3) Size selection (Ge Electrophoresis)