INTRODUCTION: Practical Uses of Genetic Engineering In this paragraph, I cover some of the practical uses of genetic engineering One

of the first an! most important "as gene splicing to help manufacture large amounts of insulin, using the cells of E coli #acteria $nother one #eing Interferon, "hich helps eliminate certain viruses an! cancer cells $nother is tissue plasminogen activator an! uro%inase, "hich are use! to !issolve #loo! clots Geneticall& engineere! #acteria is also use! to pro!uce a gro"th hormone that helps treat !"arfism'a !isor!er that ma%es &ou much, much shorter than the average human ( INTRODUCTION: DN$ E)traction *hat is DN$ E)traction goo! for+ *ell, it,s goo! for Genetic testing, -o!& i!entification, an! for anal&sis of forensic evi!ence I,ll go further into !etail "ith those in!ivi!ual sections later on DN$ E)traction: E)tracting the DN$ In or!er for &ou to #e a#le to e)tract DN$, &ou must first purif& it from proteins an! other cellular contaminants .ou !o this #& first collecting a cell, prefera#l& a chee% cell'as the& are the easiest to get(, #& s"a##ing the insi!e of the patient,s mouth, then placing the cotton s"a# in an Eppen!orf tu#e Then, ta%ing &our micropipettor, a!! some l&sis solution'The l&sis solution contains !etergent an! an en/&me calle! proteinase 0( to the tu#e This is the chemical that "ill split the cell, allo"ing &ou to get to the nucleus Place the Eppen!orf tu#e, seale!, insi!e of a "arm "ater #ath The !etergent "ill !isrupt the cell mem#rane an! the nuclear envelope, #ursting the cells open an! releasing their DN$ The DN$ "ill still #e tightl& "rappe! aroun! proteins calle! histones This is "hat the en/&me is for It cuts apart the protein, freeing the DN$ Ne)t, ta%e the tu#e out of the "arm "ater #ath, an! remove the s"a# from the tu#e 1ere, "e "ill a!! some concentrate! salt solution to the tu#e 2eal the tu#e, an! allo" the salt to clump proteins an! other !e#ris together, freeing up some space No", "e put the tu#e into a centrifuge, along "ith a tu#e of "ater on the opposite si!e to #alance it out Insi!e this centrifuge, the proteins an! !e#ris clumps "ill sin% to the #ottom, !ue to their "eight an! the rotation spee!s of the centrifuge The DN$ "ill remain free3floating in the li4ui! No", use the micropipettor to carefull& remove the top of the li4ui!'*hich contains the DN$( an! place it into a clean tu#e Then, a!! some isoprop&l alcohol to the tu#e an! invert it several times, mi)ing it into the DN$ solution Due to DN$ not #eing solu#le in isoprop&l alcohol, the DN$ "ill clump together, an! even #e visi#le to the na%e! e&e No", place the tu#e #ac% into the centrifuge an! let it run again $fter this, &ou can remove the li4ui! from the tu#e an! place it into another solution, or free/e it an! save it for another e)periment

DN$ E)traction: Genetic testing -& e)tracting the DN$ from a cell, &ou can then use the DN$ to locate several genes that &ou can compare to other genes an! fin! famil& !iseases, carrie! genetic !efaults, an! even the source of an active !efect DN$ E)traction: -o!& I!entification -& e)tracting the DN$ from human remains, "e can compare it to that of a famil& mem#er,s DN$ to i!entif& "hether or not the fore spo%en #o!& #elonge! to a mem#er of that famil& DN$ can #e gathere! from man& sources an! e)tracte!, some of these sources #eing #one marro", #iops& samples, an! the ne"#orn #a#& #loo!spot Even some of the personal items such as a hair#rush, or a tooth#rush DN$ E)traction: $nal&sis of 5orensic Evi!ence 5irst off, a sample must #e collecte! from the crime scene, an! then the DN$ #e e)tracte! using the steps I,ve liste! a#ove Then, a sample must #e collecte! from the suspects an! compare! to DN$ foun! at the crime scene, possi#l& provi!ing a match an! lan!ing &ou &our ne" convict This process inclu!es man&, man&, tests until the suspect can not #e rule! out as the source of the DN$ 2everal regions of this DN$ are copie! using a pol&merase chain reaction, or PCR PCR pro!uces millions of copies for each segment of interest an! permits minute amounts of DN$ to #e e)amine! 6ultiple regions can #e e)amine! to increase the informativeness of the test The resulting pro!ucts are then separate! an! !etecte! in or!er to characteri/e the region that is #eing e)amine! The metho!s use! to!a& inclu!e gel an! capillar& electrophoresis 5luorescence !etection metho!s greatl& ai! the sensitivit& an! ease of measuring PCR3amiplifie! 2TR alleles $fter !etecting the alleles, the num#er of repeats in a DN$ se4uence are !etermine!, a process %no"n as genot&ping *hen a result is foun!, an! is a match, the DN$ is ran through a !ata#ase to fin! the suspect, or even the victim INTRODUCTION: Gel Electrophoresis Gel Electrophoresis is use! for sorting DN$ an! proteins #& the length of their stran!s The gel use! in this process "or%s as a filter that sorts the DN$7protein stran!s using small holes .ou start #& placing &our DN$ samples into small holes at one en! of the gel Electrophoresis is ho" "e push the DN$ stran!s through the filter -& a!!ing an electrical current, "e can ma%e the DN$ move The shorter stran!s move through the gel more 4uic%l& than the longer stran!s Over time, the shorter stran!s "ill move farther a"a& from the starting point than the longer ones DN$ stran!s of the same length "ill move at the same spee! an! "in! up groupe! together, allo"ing the DN$ to sort itself 2taining the sorte! groups ma%es them visi#le to the na%e! e&e $lthough "e can,t see a single!3out stran!, "e can see larger groups The groups sho" up as #an!s in the gel

INTRODUCTION: Gel Electrophoresis In or!er to !o this ourselves, "e nee! to gather the follo"ing supplies: Po"!ere! agarose, #uffer, a flas%, a micro"ave, the gel mol! an! com# Put a small amount of agarose into a flas% an! a!! some li4ui! #uffer as "ell The #uffer is a salt "ater solution that lets electrical charges flo" through the gel 8oosel& place a plastic "rap over the top of the flas% to prevent the li4ui! from #oiling over, an! heat the mi)ture in the micro"ave until the agarose melts into the #uffer Place the mi)ture into the mol! an! place the com# into the gel on one en! Remove the com#, leaving empt& "ells for the DN$ samples No" get an electrophoresis #o) an! another #ottle of #uffer Pour the #uffer into the electrophoresis #o) Place the gel, in the mol!, into the #o) The #uffer con!ucts electrical current from one en! of the gel to the other It also %eeps the gel from !r&ing out !uring the e)periment No" &ou,ll nee! a loa!ing #uffer, DN$ sample, DN$ si/e stan!ar!, a micropipettor, the #o) containing the #uffer an! gel, an! pipette tips *ith a clean pipette tip, use the micropipettor to suc% up some loa!ing #uffer, then a!! it to the DN$ sample Ne)t, use the micropepittor to loa! the DN$ sample into the "ell No", using a clean ne" tip, use the micropepittor to suc% up some DN$ si/e stan!ar! Transfer the DN$ si/e stan!ar! into the ne)t empt& "ell The DN$ si/e stan!ar!s alrea!& has some DN$ stran!s of %no"n length, allo"ing for some eas& comparison No", hoo% up the #lac% cor! to the "ell si!e, as this has a negative charge, an! so !oes DN$ 1oo% the re! cor! into the opposite en! an! turn it on Repelle! #& the negative charge, the DN$ moves through the gel to"ar!s the positive charge at the other en! 2hort DN$ stran!s move through the holes more 4uic%l&, an! the longer ones go a #it slo"er No", "e stain the DN$ stran!s "ith a staining solution The stain is a chemical calle! ehi!ium #romi!e, "hich #in!s to DN$ an! sho"s up un!er fluorescent light No", after a#out half an hour, remove the gel an! place it on to a U9 light This "ill allo" &ou to vie" the length of the stran!s INTRODUCTION: Pol&merase Chain Reactions'PCR( PCR is a revolutionar& metho! !evelope! #& 0ar& 6ullis in the :;<=,s PCR is #ase! on using the a#ilit& of DN$ Pol&merase to s&nthesi/e a ne" stran! of DN$ complementar& to the offere! template stran! Due to Pol&merase,s limite! a#ilit& to onl& a!! a nucleoti!e to pree)isting >,3O1 groups, it re4uires a primer to "hich it can a!! the first nucleoti!e This re4uirement ma%es it possi#le to !elineate a specific region of template se4uences that the researcher "ants to amplif& -& the en! of the reaction, the specific se4uence "ill #e accumulate! in #illions of copies'amplicons ( Of course, specific en/&mes are nee!e! to successfull& s&nthesi/e ne" stran!s of DN$ One of these en/&mes #eing Ta4'from Thermis a4uaticus( DN$ pol&merase, this one #eing the most commonl& use! as "ell It has su#tle !ifferences to its si#ling en/&me, the ne)t most commonl& use!, calle! Pfu DN$ pol&merase'from p&rococcus furiosus( The things that ma%e them suita#le for PCR is, for one, the& generate ne" DN$ using a template an! primers, an! t"o, the& are #oth heat resistant

INTRODUCTION: Pol&merase Chain Reactions'PCR( The PCR reaction starts to generate copies of the template se4uence e)ponentiall&, onl& !uring this phase is it possi#le to e)trapolate #ac% to the start to !etermine the starting 4uantit& of the target se4uence containe! in the sample -ecause of the inhi#itors of the pol&merase reaction foun! in the sample, reagent limitation, accumulation of p&rophosphate molecules, an! self3annealing of the accumulating pro!uct, the reaction eventuall& ceases to s&nthesi/e the target se4uence at an e)ponential rate, causing a plateau affect This is "hat ma%es real3time 4uantitative RT3PCR so necessar& INTRODUCTION: DN$ 6icro3arra&s DN$ 6icro3arra&s are use! for e)amining thousan!s of genes at once to help locate "hich ones in!icate "hich !isor!ers The& are create! #& ro#otic machines that arrange minuscule amounts of hun!re!s of thousan!s of of gene se4uences onto a microscope sli!e *hen a gene is activate!, cellular machiner& #egins to cop& certain segments of that gene The resulting segment is %no"n as messenger RN$'mRN$( , "hich is the template for creating proteins The mRN$ pro!uce! #& the cell is complementar& an! "ill #in! to the original portion of the DN$ stran! of "hich it "as copie! $ gene is consi!ere! active if it h&#ri!i/es to the DN$ If a gene is active, it "ill sho" up as a #right fluorescent spot, revealing the possi#ilit& or certaint& of certain faults or !isor!ers