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BIOL 1000 - biotechnology - lecture notes

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BIOL 1000
Nicole Nivillac

Biology Lecture – November 14, 2013 Primer has opp ends of primes than the relication fork primes ^ Always elongated from the 3’ to 5’ Biotechnology - Bacteria is much more easier to work with since the genome is smaller and similar to ours - Heterologous – put DNA into bacteria and can be used to express proteins that aren’t theres - Bacteria have rapid growth rate - Ecoli can double in 20 mins - Fermentation technology is well established – go from small to large badges – can work with milliletres and litres and do the same job - Cannot do glycolysis – do not have golgi - Cannot remove introns unlike our DNA (introns are removed) Cloning - Recombinant DNA – DNA from different organisms is combined into one organism - Polymerase chain reaction – takes DNA from breaking a cell open and copies is a lot of times - ^ template DNA, primer, polymerase, and nucleotides are needed for this reaction - Denaturation – heating temperature so that strands are separated - Bonds between As and Ts will break since they have more bonds to be broken - Anneal – cools it down so primers can bind (primers are designed since DNA sequence are known) - Extend – polymerase elongates the DNA strands - Too many enzymes can hinder the process ^ reason why helicase and other stuff is not added - Introns are not removed because RNA primers are not used Agarose Gel Electrophoresis - Made of Agarose substances - DNA moves from (-) to (+) – DNA is negatively charged so it moves to the (+) charge - Bigger fragments don’t move fast whereas smaller ones move fast - Produces PCR results - To visualize the DNA, florescent dye is added to illuminate the DNA so that it can be seen under UV light - After amplifying the DNA, digestion happens so that certain DNA can be cut and be used - ^ cuts at restriction sites (palindromic) - ^ same sequence from both sides EX. (5’ GAATTC 3’ same as 3’ CTTAAG 5’) - Sticky ends contain uncomplemented bases whereas blunt ends don’t - MCS – multiple cloning site is the place where the bacterial plasmid DNA be digested - ^ where the plasmid is cut (digested) – sticky ends are pr
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