Date post: | 04-Apr-2018 |
Category: |
Documents |
Upload: | stevin-wilson |
View: | 224 times |
Download: | 0 times |
of 58
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
1/58
Rmlow Imfndo Jhf~rh gkr Bok~hjdfkakiy
Nhpmr~chf~ kg Bok~hjdfkakiy! Cofoz~ry kg Zjohfjh ' ^hjdfkakiy! ^dorxwmfmf~dmpxrmc! Ehrmam
ZFP Ihfk~ypofi mfn Jhaa Jxa~xrh ^hjdfo}xhz
Dxcmf Ckahjxamr Ihfh~ojz Amb
Zxbco~~hn b
Z~hwof Yoazkf
Mrxfm Ckdmf
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
2/58
2
Mjefkyahnihchf~Yh ykxan aoeh ~k htprhzz kxr irm~o~xnh ~k maa ~dkzh ydk imwh xz ~dh pkzzoboao~ ~k
jkcpah~h ~doz prklhj~&
Gorz~ kg maa yh ykxan aoeh ~k ~dmfe Nr&Rmndmerozdfmf Poaamo! Norhj~kr! Rmlow Imfndo Jhf~rh
gkr Bok~hjdfkaki mfn Rhzhmrjd gkr iowofi xz ~dh kppkr~xfo~ ~k bh m pmr~ kg ~dh ofz~o~x~h&
Kxr dhmr~gha~ ~dmfez ~k Nr& Ckofme Bmfhrlhh gkr ah~~ofi xz xzh doz amb! gkr doz kx~z~mfnofi
zxppkr~ mfn efkyahnih dh zdmrhn yo~d xz&
^doz rhpkr~ ykxan fk~ dmwh bhhf pkzzobah yo~dkx~ ~dh dhap kg Cz& Zym~d B! Cr&Zmfozd
Zm~dmf! Cz& Zmrmnm Amezdco!Cr& Zdmbhhzd Bmamf! Cr& Mf~kf EP mfn Cz& Mfmzymrm
Mzdke& ^dh z~hhrhn xz ~drkxid ~dh bmzojz kg maa ~hjdfo}xhz prmj~ojhn of ~dh amb& Yh
ykxan aoeh ~k ~dmfe Crz& Whax~dmo gkr prkwonofi xz gmjoao~ohz of ~dh amb&
Yh mazk mpprhjom~h ~dh zxppkr~ mfn hfjkxrmihchf~ kg kxr ghaaky amb cm~hz nxrofi ~dh
~hfxrh&
Mbkwh maa yh ~dmfe ~dh Macoid~ gkr Doz bahzzofiz yo~dkx~ ydojd ~doz ~rmofofi ykxan dmwh
bhhf lxz~ m nrhmc&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
3/58
:
Jkf~hf~z
Zhj~okf#M< ZFP Ihfk~ypofi
M=&NFMOZKAM^OKF7
M2&NFM]XMF^OGOJM^OKFMFN]XMAO^_JDHJEKGNFM =0
M:&PKA_CHRMZHJDMOFRHMJ^OKF =:
M>&MIMRKZHIHAHAHJ^RKPDKRHZOZ=?
M8&NFMZH]XHFJOFI 2=
M7&RHZ^ROJ^OKFGRMICHF^AHFI^DPKA_CKRPDOZC 29
^MBAHZMFNGOIXRHZ:0
B=&PRHPMROFIMFMZHP^OJHFWORKFCHF^:>
B2&PRHPMRM^OKFKGJHAAIRKY^DCHNOXCMFNJXA^XRHJKFNO^OKFZ:7
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
4/58
>
B:&JDHJEOFIJHAAZ:?
B>&ZXBJXA^XRHGRKCMNDHRHF^JXA^XRHNJHAAZ>0
B8&JKXF^OFIJHAAZXZOFIDHCKJ_^KCH^HR>2
B7&GRHH\OFIJHAAZ>:
B9&JHAARHWOWMA>8
B4&JHAAZHHNOFIGKRNFM+RFMOZKAM^OKF>9
B?&NRXI^RHM^CHF^^KJHAAAOFH>4
B=0&NFMOZKAM^OKFGRKCJXA^XRHNJHAAAOFHZ>?
B==&]XMF^OGOJM^OKFXZOFIFMFKNRKP80
B=2&RFMOZKAM^OKFGRKCJXA^XRHNJHAAAOFHZ8=
B=:&MZZHZZCHF^KGRFM]XMAO^_MFN]XMF^O^_82
B=>&jNFMPRHPMRM^OKF8:
B=8&RHMA#^OCHPJR88
RHGHRHFJHZ84
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
5/58
8
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
6/58
7
Htphrochf~ #M=
NFM OZKAM^OKF
Bhgkrh mf ~ph kg mfmazoz jmf bh phrgkrchn! NFM cxz~ bh ozkam~hn&
^dh zxjjhzz kg maa zxbzh}xhf~ prkjhnxrhz nhphfnz kf ~dh mwmoamboao~ kg
zxggojohf~ mckxf~z kg NFM kg ~dh mpprkprom~h }xmao~& Cmf nogghrhf~ ch~dknz
mfn ~hjdfkakiohz mrh mwmoambah gkr ~dh ozkam~okf kg ihfkcoj NFM& Maa ch~dknz
ofwkawh nozrxp~okf mfn azoz kg ~dh z~mr~ofi cm~hroma gkaakyhn b ~dh rhckwma kg
prk~hofz mfn k~dhr jkf~mcofmf~z mfn gofmaa rhjkwhr kg ~dh NFM&
^dh ch~dkn xzhn dhrh oz Pdhfka jdakrkgkrc NFM ht~rmj~okf grkc bakkn
zmcpah& ^doz ch~dkn ymz gkxfn b ymz kroiofmaa nhwozhn b Pok~r Jdkcj|fzeo
mfn Fojkah~~m Zmjjdo mfn pxbaozdhn of =?49 $rhghrrhn ~k mz Ixmfonofoxc
~dokjmfm~h#pdhfka#jdakrkgkrc ht~rmj~okf/&
Profjopahj&
Jkaahj~ ~dh zxphrfm~mf~ of~k m grhzd ~xbh mfn mnn 8ca kg ^roz Pdhfka& Cot
~dh jkf~hf~z kg ~dh ~xbh ihf~a gkr 2cofx~hz mfn ~dhf mnn 8ca kg
Jdakrkgkrc% ozkmca majkdka $28j kr #20j kr#40j gkr gx~xrh xzh&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
10/58
=0
Htphrochf~#M2
NFM ]XMF^O^M^OKF MFN ]XMAO^_ JDHJE KG NFM
Prokr ~k mf mfmazoz! NFM zmcpahz zdkxan bh }xmf~o~m~hn mfn jdhjehngkr pxro~ kg NFM& ^dh mckxf~ kg aoid~ ~dm~ m zmcpah mbzkrbz m~ m pmr~ojxamr
ymwhahfi~d oz chmzxrhn mfn xzhn ~k nh~hrcofh ~dh jkfjhf~rm~okf kg ~dh zmcpah b
jkcpmrozkf yo~d mpprkprom~h z~mfnmrnz kr rhghrhfjh nm~m& ^dh ckz~ jkcckfa
xzhn ch~dknkakiohz gkr }xmf~ogofi ~dh mckxf~ kg fxjahoj mjon of m prhpmrm~okf
mrh< $o/ iha hahj~rkpdkrhzoz mfn $oo/ zphj~rkpdk~kch~roj mfmazoz& Dhrh ~dh zmcpah
mckxf~ bhofi ahzz! ~dh am~~hr ch~dkn oz prhghrrhn&
m/ Zphj~rkpdk~kch~roj Nh~hrcofm~okf
^dh zphj~rkpdk~kch~roj }xmf~ogojm~okf kg NFM oz bmzhn kf Bhhr#Amcbhr~z amy ~dm~
iowhz ~dh aofhmr rham~okfzdop bh~yhhf mbzkrbmfjh mfn jkfjhf~rm~okf kg mbzkrbofi
zphjohz& Whrog ~dh kb~mofhn prochr b hf~hrofi of~k XJZJ&
8& Ofpx~ ~dh kb~mofhn prochr of~k Prochr Prhcohr ~k jdhje og o~ gkrcz dmorpof
z~rxj~xrh&
Irmnohf~ PJR
^dh zhahj~okf kg ~dh mffhmaofi ~hcphrm~xrh oz pkzzoba ~dh ckz~ jro~ojma
jkcpkfhf~ gkr kp~oco|ofi ~dh zphjogojo~ kg m PJR rhmj~okf& Of ckz~ jmzhz! ~doz
~hcphrm~xrh cxz~ bh hcporojmaa ~hz~hn& ^dh PJR oz fkrcmaa z~mr~hn m~ 8J
bhaky ~dh jmajxam~hn ~hcphrm~xrh kg ~dh prochr cha~ofi pkof~ $^c/& Dkyhwhr!
~dh pkzzobah gkrcm~okf kg xfzphjogoj zhjkfnmr bmfnz zdkyz ~dm~ ~dh kp~ocxc
~hcphrm~xrh oz kg~hf cxjd doidhr ~dmf ~dh jmajxam~hn ~hcphrm~xrh $6=2J/&
Gxr~dhr PJR rhmj~okfz yo~d irmnxmaa ofjrhmzofi ~hcphrm~xrhz mrh rh}xorhn
xf~oa ~dh ckz~ z~rofihf~ jkfno~okfz dmwh bhhf gkxfn& Ydhf m z~mfnmrn PJR
jjahr oz xzhn! ~doz ch~dkn oz ~dh ckz~ ~och#of~hfzowh kp~oco|m~okf z~rm~hi&
Nxrofi ~dh PJR! m ~hcphrm~xrh irmnohf~! ydojd jmf bh prkirmcchn bh~yhhfzm 80 mfn 7>J! oz bxoa~ xp mjrkzz ~dh ~dhrck bakje& ^doz maakyz ~dh ckz~
z~rofihf~ pmrmch~hrz gkr hwhr prochr zh~ ~k bh jmajxam~hn yo~d ~dh mon kg kfa
kfh zofiah PJR rhmj~okf&
^dh gkaakyofi rhmj~okf cot~xrh ymz prhpmrhn gkr m rh}xorhn fxcbhr kg
rhmj~okfz
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
16/58
=7
JKCPKFHF^Z WKAXCH$a/
NFM
=0T Bxgghr
nF^Pz
20 pC gkrymrn prochr
20 pC rhwhrzh prochr
^m} pkachrmzh $:X/
Z~hroah ym~hr
=
=
=
0&=
0&=
0&=2
7&74
^k~ma =0
Wkr~ht ~dh cot~xrh brohga! ~dhf jhf~rogxih m~ aky zphhn& ^dh Irmnohf~ PJR ymz
phrgkrchn yo~d ~dh mbkwh ~xbhz of m ~dhrcma jjah mjjkrnofi ~k ~dh gkaakyofi
prk~kjka
Ofo~oma nhfm~xrm~okf
Nhfm~xrm~okf
Mffhmaofi
Ht~hfzokf
Gofma ht~hfzokf
Dkan
?>J
?>J
880J ~k 78kJ
92J
92J
>kJ
8 cofx~hz
:0zhjkfnz
:0zhjkfnz
:0zhjkfnz
8 cofx~hz
Gkrhwhr
Prk~kjka gkr m PJR rhmj~okf
=&^dh htphrochf~ dmz ~k bh pamffhn prokr ~k mf mnno~okf kg rhmihf~z
$Fxcbhr kg prochr pmorz ~k bh xzhn! fxcbhr kg NFM ~hcpam~hz! h~j&/&
Rhmihf~z ]xmf~o~ phr PJR ~xbh of a
Noz~oaahn ym~hr 7&74
=0t Bxgghr =&0
2&8 C nf~p =&0
20pC Gkrymrn Prochr 0&=
:8Jyjahz
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
17/58
=9
2& Mg~hr nkofi zk! cmeh ~dh mpprkprom~h jkje~moa+z mfn hfzxrh jkcpah~h
cotofi b ~mppofi ~dh ~xbh mfn }xoje zpoffofi&
$F&B& Jmx~okf zdkxan bh xzhn ~k mwkon jkf~mcofm~okf kg rhmj~okfz yo~d hwhf
zcmaa mckxf~z kg NFM& Of mnno~okf! jmrh zdkxan bh ~mehf ~k mwkon
jkf~mcofm~okf kg poph~~h yo~d jmrrkwhr mcpaogojm~okf prknxj~z grkc
prhwokxz rhmj~okfz/
:& Poph~~h ?&: a kg ~dh mpprkprom~h jkje~moa norhj~a of~k ~dh bk~~kc kg m
z~hroah cojrkhpphfnkrg ~xbh gkr hmjd rhmj~okf& ^dh ~xbhz zdkxan bh
ambhahn&
>& Mnn 0&9 a kg ~dh NFM norhj~a of~k ~dh nrkp kg jkje~moa of hmjd ~xbh mfn
hfzxrh mnh}xm~h cotofi& ]xoje zpof ~k jkaahj~ ~dh rhmj~okf cot~xrh of ~dh
bk~~kc kg ~dh ~xbh&
8& Pamjh ~dh ~oid~a jmpphn ~xbhz of ~dh ~hcphrm~xrh bakje mfn cmeh zxrh
hmjd oz gorca zhm~hn b prhzzofi kf ~dh ~xbhz ofnowonxmaa&
^dh PJR cmjdofh cxz~ fky bh prkirmcchn gkr ~dh zphjogoj rhmj~okf
jkfno~okfz nhzorhn& Hmjd jjah of ~dh pkachrmzh jdmof rhmj~okf ofwkawhz
~drhh z~hpz $nhfm~xrofi! prochr mffhmaofi! pkachro|m~okf/! mfn ~dh
prknxj~z mrh mcpaogohn b phrgkrcofi cmf jjahz kfh mg~hr ~dh k~dhr
yo~d ~dh dhap kg ~dh mx~kcm~hn ~dhrcma jjahr&
^dh ^m} pkachrmzh oz dhm~ z~mbah! mfn rhcmofz mj~owh nhzpo~h ~dh doid
nhfm~xrofi ~hcphrm~xrh kg hmjd jjah& M rhprhzhf~m~owh zh~ kg rhmj~okfjkfno~okfz gkr 28#:8 jjahz ozJ
?>J
880J
92J
92J
>kJ
8 cofx~hz
:0zhjkfnz
:0zhjkfnz
:0zhjkfnz
8 cofx~hz
Gkrhwhr
7& Mg~hr jkcpah~okf kg ~dh PJR rhmj~okf! rhckwh ~dh ~xbhz grkc ~dh
~hcphrm~xrh bakje mfn pamjh ~dhc of mf hpphfnkrg rmje&
9&^dh rhmj~okf prknxj~z mrh jkfwhfohf~a zhpmrm~hn mjjkrnofi ~k zo|h b
hahj~rkpdkrhzoz ~drkxid m =( pkamjramconh iha m~ 98 W gkr :0#
>8cofx~hz! mfn wozxmao|hn mg~hr z~mofofi ~dh iha yo~d h~donoxc brkconh&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
19/58
=?
Htphrochf~#M>
IHA HAHJ^RKPDKRHZOZ
O~ oz xzhn ~k zhpmrm~h NFM grmichf~z& Hahj~rkpdkrhzoz xzhz mf hahj~roj jxrrhf~
~k zhpmrm~h nogghrhf~#zo|hn ckahjxahz of m pkrkxz! zpkfih#aoeh cm~rot& Zcmaahr
ckahjxahz ckwh ckrh hmzoa ~drkxid ~dh iha pkrhz ~dmf amrihr ckahjxahz&
^dh ~hjdfo}xh xzhz mf mimrkzh iha! cmnh grkc doida pxrogohn zhmyhhn& ^doz
jkxan bh xzhn ~k zhpmrm~h NFM ckahjxahz rmfiofi grkc zhwhrma dxfnrhn
fxjahk~onhz of ahfi~d ~k kwhr =0!000 fxjahk~onhz&
Cm~hromaz rh}xorhn
Dkro|kf~ma iha hahj~rkpdkrhzoz
Iha ~rm
Iha jkcbz
Pkyhr zxppa xfo~
Cojrk ymwh kwhf
XW ~rmfzoaaxcofm~kr
Rhmihf~z rh}xorhn
=T ^MH bxgghr
H~Br $ H~donoxc Brkconh/
Iha akmnofi nh$krmfih I/
Mimrkzh $xa~rm pxrh/
Prkjhnxrh
=&^dh iha oz prhpmrhn b cha~ofi 0&:i kg mimrkzh of :0ca kg =T ^MH bxgghr&
Mnn 2 a kg H~donoxc brkconh of~k ~dh cot~xrh mfn ~dh cot ymz pkxrhn
of~k ~dh iha ~rm ~mphn kf maa zonhz&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
20/58
20
2&^dh jkcbz mrh pamjhn of ~dh zak~z mfn ~dh iha oz rhmn ~k bh xzhn! kfjh o~
zh~z&
:&^dh ~mph oz rhckwhn mfn ~dh iha oz zxbchrihn of m ~mfe goaahn yo~d =T^MH
bxgghr ~dm~ jkfnxj~z hahj~rojo~&
>& Xzofi m poph~~h! NFM zmcpahz mrh akmnhn of~k ~dh yhaaz cmnh of ~dh
mimrkzh iha& ^dh NFM zmcpahz mrh jkakxrahzz! bx~ m baxh ~rmjeofi nh oz
mnnhn ~k ~rmje ~dh NFM coirm~okf ~drkxid ~dh iha&
8&^dh pdkzpdm~h irkxpz of ~dh NFM bmjebkfh jmrr fhim~owha jdmrihn
ktihf iowofi m NFM ckahjxah mf kwhrmaa fhim~owh jdmrih& Of m f hahj~roj
jxrrhf~! ~dh fhim~owha jdmrihn NFM ckwhz ~kymrn ~dh pkzo~owh pkah kg
~dh hahj~rkpdkrhzoz jdmcbhr&
7&^dh NFM ckahjxahz ckwh ~drkxid ~dh iha b rhpx~m~okf# m rhp~oah#aoeh
zfmeofi mj~okf ~drkxid ~dh pkrhz kg ~dh mimrkzh cm~rot& Zcmaahr NFM
grmichf~z coirm~h gmz~hr mfn gxr~dhr kwhr m iowhf phrokn kg ~och ~dmf nk
amrihr grmichf~z& ^doz oz dky NFM grmichf~z jmf bh zhpmrm~hn b zo|h of
m mimrkzh iha&
9& M pdk~k kg ~dh iha oz ~mehf gkr am~hr mfmazoz&
4&^dh zo|h kg mf NFM grmichf~ jmf bh nh~hrcofhn b jkcpmrofi o~ ~k
cmrehrz# NFM grmichf~z kg efkyf zo|hz&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
21/58
2=
Htphrochf~#M8
NFM ZH]XHFJOFI
Profjopah
^dh profjopahz kg NFM rhpaojm~okf yhrh xzhn b Zmfihr h~ ma& $=?9>/ of ~dh
nhwhakpchf~ kg ~dh prkjhzz fky efkyf mz Zmfihr nonhkt zh}xhfjofi& ^doz
prkjhzz ~mehz mnwmf~mih kg ~dh mboao~ kg NFM pkachrmzh ~k ofjkrpkrm~h 2 ! :#
nonhktfxjahk~onhz! fxjahk~onh bmzh mfmakiz ~dm~ amje ~dh :!#dnrkta irkxp
hzzhf~oma of pdkzpdknohz~hr bkfn gkrcm~okf& Zmfihr nonhkt zh}xhfjofi
rh}xorhz m NFM ~hcpam~h! m zh}xhfjofi prochr! NFM pkachrmzh! fxjahk~onhz
$nF^Pz/! nonhktfxjahk~onhz $nnF^Pz/! mfn rhmj~okf bxgghr&
Gkxr zhpmrm~h rhmj~okfz mrh zh~ xp! hmjd jkf~mofofi rmnokmj~owha ambhahn
fxjahk~onhz mfn ho~dhr nnM! nnJ! nnI! kr nn^& ^dh mffhmaofi! ambhaofi! mfn
~hrcofm~okf z~hpz mrh phrgkrchn kf zhpmrm~h dhm~ bakjez& NFM zf~dhzoz oz
phrgkrchn m~ :9 J! ~dh ~hcphrm~xrh m~ ydojd ~dh ^9 NFM pkachrmzh xzhn dmz
~dh kp~ocma hf|ch mj~owo~&
NFM pkachrmzh mnnz ho~dhr m nhktfxjahk~onh kr ~dh jkrrhzpkfnofi 2 ! :#
nonhktfxjahk~onh m~ hmjd z~hp kg jdmof ht~hfzokf& Ydh~dhr m nhktfxjahk~onh
kr m nonhktfxjahk~onh oz mnnhn nhphfnz kf ~dh rham~owh jkfjhf~rm~okf kg bk~d
ckahjxahz& Ydhf m nhktfxjahk~onh $M! J! I! kr ^/ oz mnnhn ~k ~dh : hfn! jdmof
ht~hfzokf jmf jkf~ofxh& Dkyhwhr! ydhf m nonhktfxjahk~onh $nnM! nnJ! nnI! kr
nn^/ oz mnnhn ~k
~dh : hfn! jdmof ht~hfzokf ~hrcofm~hz & Zmfihr nonhkt zh}xhfjofi rhzxa~z of
~dh gkrcm~okf kg ht~hfzokf prknxj~z kg wmrokxz ahfi~dz ~hrcofm~hn yo~d
nonhktfxjahk~onhz m~ ~dh : hfn&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
22/58
22
NFM ^hcpam~h Prhpmrm~okfJ
=0 zhj
> cof
Gkrhwhr
b/ Zh}xhfjofi Rhmj~okf Jahmf Xp
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
25/58
28
nF^Pz! hf|chz! zdkr~! gmoahn PJR prknxj~ zk ~dm~ ~dh nk fk~ of~hrghrh yo~d
~dh nkyfz~rhmc mppaojm~okf zxjd mz jakfofi zh}xhfjofi kr ambhaofi&
Cm~hromaz mfn rhmihf~z?
Dh~hrk|ikxz JI 2>?!=:>!==8
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
32/58
:2
Goixrh >
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
33/58
::
ZHJ^OKF#B
MFOCMA JHAA JXA^XRH
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
34/58
:>
Htphrochf~ B=
PRHPMROFI MF MZHP^OJ JKFNO^OKF
Moc^k hfzxrh jhaa jxa~xrh prkjhnxrhz mrh phrgkrchn ~k m z~mfnmrn ~dm~ yoaa prhwhf~
jkf~mcofm~okf grkc bmj~hrom! gxfio!mfn cjkpamzcm mfn jrkzz jkf~mcofm~okf
yo~d k~dhr jhaa aofhz&
Cm~hromaz rh}xorhn
90( h~dmfka of ym~hr
H}xopchf~
Phrzkfma prk~hj~owh h}xopchf~ $z~hroah iakwhz! ambkrm~kr jkm~/ Cojrkbokakiojma zmgh~ jmbofh~ m~ mpprkprom~h jkf~mofchf~ ahwha
Prkjhnxrh
=& Zmfo~o|h ~dh jmbofh~ xzofi 90( h~dmfka bhgkrh jkcchfjofi ykre&
2& Zmfo~o|h iakwhz b ymzdofi ~dhc of 90( h~dmfka mfn maakyofi nrofi gkr :0
zhjkfnz bhgkrh jkcchfjofi ykre&
:& Px~ maa cm~hromaz mfn h}xopchf~ of~k ~dh jmbofh~ prokr ~k z~mr~ofi ykre mg~hr
zmfo~o|ofi ~dh ht~hrokr zxrgmjhz yo~d 90( h~dmfka&
>& Nozjmrn iakwhz mg~hr dmfnaofi jkf~mcofm~h jxa~xrhz mfn m~ ~dh hfn kg maa jhaa
jxa~xrh prkjhnxrhz&
8& Ckwhchf~ yo~dof mfn occhnom~ha kx~zonh ~dh jmbofh~ cxz~ fk~ bh rmpon&
Zaky ckwhchf~ yoaa maaky ~dh mor yo~dof ~dh jmbofh~ ~k jorjxam~h prkphra
7& Zphhjd!zfhh|ofi mfn jkxidofi cxz~ bh norhj~hn mym grkc ~dh jmbofh~ zk mz
fk~ ~k nozrxp~ ~dh morgaky&
9& Mg~hr jkcpah~ofi ykre nozofghj~ maa h}xopchf~ mfn cm~hromaz bhgkrh rhckwofi
grkc ~dh jmbofh~& Zprm ~dh ykre zxrgmjhz ofzonh ~dh jmbofh~ yo~d 90(
h~dmfka mfn yoph yo~d ~ozzxh pmphr&
4&Phroknojmaa jahmf ~dh jmbofh~ zxrgmjhz yo~d m nozofghj~mf~
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
35/58
:8
Ch~dkn gkr jahmfofi JK2 ofjxbm~kr mfn bokzmgh~ jmbofh~
4&m&Jahmf JK2 ofjxbm~kr yo~d 2&8( zknoxc dpkjdakro~h
4 b& Ahmwh gkr 8 cof& Rofzh yo~d ym~hr mfn rhckwh ym~hr jkcpah~ha xzofi
~ozzxh&
4&j&Zprm ofjxbm~kr yo~d 90( Ozkprkpmfka& Yoph yo~d nr ~ozzxh ~k rhckwh mf
rhzonxma zknoxc dpkjdakro~h mfn ym~hr&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
36/58
:7
Htphrochf~ B2
PRHPMRM^OKF KG JHAAIRKY^D CHNOXC MFN JHAA IRKY^D
JKFNO^OKFZ
Moc
Bhgkrh z~mr~ofi ykre jdhje ~dh ofgkrcm~okf iowhf yo~d ~dh jhaa aofh ~k onhf~og
ydm~ chnom ~ph! mnno~owhz mfn rhjkcchfnm~okfz zdkxan bh xzhn&
Ckz~ jhaa aofhz jmf bh irkyf xzofi NCHC jxa~xrh chnom kr RPCO jxa~xrh chnom
yo~d =0( Gh~ma Bkwofh Zhrxc $GBZ/ mfn mf~obok~ojz jmf bh mnnhn og rh}xorhn&
Ckz~ jhaa aofhz yoaa irky kf jxa~xrh gamzez yo~dkx~ ~dh fhhn gkr zphjoma cm~rothz
h~j& Dkyhwhr! zkch jhaaz! pmr~ojxamra procmr jhaaz! yoaa rh}xorh irky~d kf
zphjoma cm~rothz zxjd mz jkaamihf ~k prkck~h jhaa m~~mjdchf~! nogghrhf~om~okf kr
jhaa irky~d&
Cm~hromaz rh}xorhn
=&NCHC
2&GBZ:&Mf~ocjk~oj mf~obok~oj zkax~okf
Prkjhnxrh
m/ Chnom prhpmrm~okfo& Prhpmrm~okf kg NCHC
M& Mnn pkynhrhn chnoxc ~k =8J ~k :0 J $Rkkc ~hcphrm~xrh/
mx~kjamwhn ym~hr yo~d ihf~ah z~orrofi&$Nk fk~ dhm~/&
B& Rofzh kx~ ofzonh kg pmjeh~ ~k rhckwh ~rmjhz kg pkynhr&
J& Mnn =&9i kg FmDJK: phr ao~rh kg chnoxc&
N&Noax~h ~k nhzorhn wkaxch ym~hr& Z~or xf~oa nozzkawhn&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
37/58
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
38/58
:4
=&NCHC >8ca
2&=0( GBZ 8 ca
:&Mf~obok~oj mf~ocjk~oj zkax~okf 0&8 ca
80 ca
Prkwonofi jxa~xrofi jkfno~okfz
Jhaa aofhz mrh cmof~mofhn m~ :90J ofjxbm~kr m~ 8( JK2&
Gkr dxcmf jhaaz! jkm~ gamzez yo~d =( iham~of&
Prhpmrh =0cA kg jkm~ofi zkax~okf jkcpkzhn kg =( iham~of b noax~ofi yo~d
noz~oaahn ym~hr! gkaakyhn b goa~rm~okf& ^doz oz hggojohf~ ~k jkm~ mbkx~ 8 gamzez&
=& Poph~~h jkm~ofi zkax~okf of~k gamze& Rkje bmje mfn gkr~d ~k hwhfa
noz~robx~h ~dh bk~~kc kg ~dh gamze& Ah~ zo~ of mf ofjxbm~kr gkr =8#:0
cofx~hz&
2& Jkcpah~ha rhckwh jkm~ofi zkax~okf b mzporm~ofi bhgkrh zhhnofi&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
39/58
:?
Htphrochf~ B:JDHJEOFI JHAAZ
Jhaaz zdkxan bh jdhjehn cojrkzjkpojmaa nmoa ~k hfzxrh ~dh mrh dhma~d
mfn irkyofi mz htphj~hn& M~~mjdhn jhaaz zdkxan bh cmofa m~~mjdhn ~k ~dh
bk~~kc kg ~dh gamze! rkxfn mfn paxcp kr hakfim~hn of zdmph mfn rhgrmj~ofi aoid~
mrkxfn ~dhor chcbrmfh& Zxzphfzokf jhaaz zdkxan akke rkxfn mfn paxcp mfn
rhgrmj~ofi aoid~ mrkxfn ~dhor chcbrmfh& Zkch zxzphfzokf jhaaz cm jaxcp&
Chnom zdkxan bh pofe#krmfih of jkakr&
Nozjmrn jhaaz og0
Htphrochf~ B>ZXBJXA^XRH KG MNDHRHF^ JHAA AOFHZ
Moc
Mndhrhf~ jhaa aofhz yoaa irky of wo~rk xf~oa ~dh dmwh jkwhrhn ~dh zxrgmjh mrhm
mwmoambah kr ~dh chnoxc oz nhpah~hn kg fx~rohf~z& M~ ~doz pkof~ ~dh jhaa aofhz
zdkxan bh zxb#jxa~xrhn of krnhr ~k prhwhf~ ~dh jxa~xrh nofi& Jhaa pmzzmiofi kr
zpao~~ofi oz m ~hjdfo}xh ~dm~ hfmbahz mf ofnowonxma ~k ehhp jhaaz maowh mfn
irkyofi xfnhr jxa~xrhn jkfno~okfz gkr ht~hfnhn phroknz kg ~och& Jhaaz zdkxanbh pmzzhn ydhf ~dh mrh 48(#?0( jkfgaxhf~& ^k zxbjxa~xrh ~dh jhaaz ~dh fhhn
~k bh brkxid~ of~k zxzphfzokf& ^dh nhirhh kg mndhzokf wmrohz grkc jhaa aofh ~k
jhaa aofh bx~ cmlkro~ kg jmzhz prk~hmzhz! h&i& ~rpzof !mrh xzhn ~k rhahmzh ~dh
jhaaz grkc ~dh gamze! Dkyhwhr! ~doz cm fk~ bh mpprkprom~h gkr zkch ~och ydhrh
htpkzxrh ~k prk~hmzhz oz dmrcgxa kr ydhrh ~dh hf|chz mrh xzhn ~k rhckwh
chcbrmfh cmrehrz+rhjhp~krz kg of~hrhz~& Of ~dhzh jmzhz jhaaz zdkxan bh
brkxid~ of~k zxzphfzokf of~k m zcmaa wkaxch kg chnoxc chjdmfojmaa yo~d ~dh
mon kg jhaa zjrmpphrz&
Cm~hromaz rh}xorhn
NCHC#GBZ chnoxc
PBZ+HN^M zkax~okf
0&08( ^rpzof
H}xopchf~
Phrzkfma prk~hj~owh h}xopchf~
Cojrkbokakiojma zmgh~ jmbofh~
JK2 ofjxbm~kr
Prh#ambhahn gamzez
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
41/58
>=
Cmrehr phf
Cojrk poph~~hz
Mcpxah rmje
Prkjhnxrh
=& Nozjmrn ~dh chnom grkc ~dh jxa~xrh gamze&
2& Ymzd ~dh jxa~xrh gamze ~yojh yo~d 2ca PBZ+HN^M&
:& Mnn ^rpzof mfn ihf~a zdmeh o~ zk ~dm~ jhaaz ih~ nh~mjdhn&
>& Mnn =ca grhzd chnom ~k nhmj~owm~h ~rpzof
8&^rmfzghr ~k m jhf~rogxih ~xbh
7& Jhf~rogxih m~ =000rpc gkr : cof
9& Nozjmrn zxphrfm~mf~
4& Mnn =ca grhzd chnom mfn ~rmfzghr ~k jxa~xrh gamze&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
42/58
>2
Htphrochf~ B8
JKXF^OFI JHAAZ YO^D DHCKJ_^KCH^HR
m& Prhpmrofi dhckj~kch~hr
o& Hfzxrh ~dh dhckj~kch~hr oz jahmf xzofi 90( h~dmfka&
oo& Ckoz~hf ~dh zdkxanhrz kg ~dh dhckj~kch~hr mfn mggot ~dh jkwhrzaop xzofi
ihf~ah prhzzxrh mfn zcmaa jorjxamr ck~okfz& ^dh pdhfkchfkf kg Fhy~kfz rofiz
jmf bh kbzhrwhn ydhf ~dh jkwhrzaop oz jkrrhj~a mggothn! ~dxz ~dh nhp~d kg ~dh
jdmcbhr oz hfzxrhn&
b& Jkxf~ofi
o& Xzofi ~dh Ioazkf poph~~h! nrmy xp zkch jhaa zxzphfzokf jkf~mofofi ~rpmf
baxh& Jmrhgxaa goaa ~dh dmhckj~kch~hr b ihf~a rhz~ofi ~dh hfn kg ~dh Ioazkf
~op m~ ~dh hnih kg ~dh jdmcbhrz& ^meh jmrh fk~ ~k kwhr# goaa ~dh jdmcbhr& Maaky
~dh zmcpah ~k bh nrmyf kx~ kg ~dh poph~~h b jmpoaamr mj~okf! ~dh gaxon zdkxan
rxf ~k ~dh hnihz kg ~dh irkkwhz kfa& Rh#akmn ~dh poph~~h mfn goaa ~dh zhjkfn
jdmcbhr og rh}xorhn&
oo& Gkjxz kf ~dh iron aofhz kg ~dh dhckj~kch~hr xzofi ~dh =0T kblhj~owh kg ~dhcojrkzjkph& Gkjxz kf kfh zh~ kg =7 jkrfhr z}xmrh m~ ~dh jkrfhrz&
ooo& ^meh ~dh mwhrmih kg ~dh fxcbhr kg jhaaz gkxfn m~ ~dh jkrfhrz&
ow& Jhaaz phr ca 3 ~dh mwhrmih jkxf~ phr z}xmrh t ~dh noax~okf gmj~kr t =0>
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
43/58
>:
Htphrochf~#B7
GRHH\OFI KG JHAA#AOFH
Moc
O~ oz jkcckf prmj~ojh ~k jrhm~h m cmz~hr bmfe jkfzoz~ofi kg 2 ~k 20 womaz kg ~dh
jhaa aofh& ^dhf jrhm~h kfh kr ~yk ykreofi bmfez grkc ~doz yo~d 2 ~k 20 womaz of
hmjd $nhphfnofi kf dky kg~hf ~dh jhaaz yoaa bh rh}xorhn/& Ydhf ~dh ykreofi
bmfe oz xzhn xp! m fhy ykreofi bmfe jmf bh jxa~xrhn mfn jrhm~hn grkc kfh
woma kg ~dh kroiofma cmz~hr bmfe& Og pkzzobah! ehhp ~dh cmz~hr mfn ykreofi bmfe
of zhpmrm~h ao}xon fo~rkihf z~krmih ~mfez&
^doz yoaa hfzxrh kx maymz dmwh m z~kje kg jhaaz grkc m akyhr pmzzmih fxcbhr
mfn o~ yoaa mazk fk~ bh fhjhzzmr ~k ehhp pxrjdmzofi ~dh jhaa aofh&
Cm~hromaz
=& = ca # 2 ca jrkwoma
2& Jhaa jxa~xrh chnoxc yo~d 20( GBZ $Gh~ma bkwofh zhrxc/ mfn fhjhzzmr
zxppahchf~z
:& NCZK $Noch~da zxagktonh/! doid pxro~! z~hroah! gkr jhaa jxa~xrh
>& Prhpmrh grhh|ofi chnoxc< ~k jhaa jxa~xrh chnoxc! mnn 8#=0( $w+w/
NCZK&
Prkjhnxrh
=& Zpao~ ~dh jhaaz& ^meh =ca ~k jrkwoma&2& Jhf~rogxih m~ 2800rpc gkr : cofx~hz m~ > J&
:& Nozjmrn ~dh zxphrfm~mf~ mfn mnn 980xa grhh|ofi cot~xrh $?ca GBZ % =ca
NCZK/&
>& Z~krh m~ #20 J gkr = nm! ~dhf of #90 J $2#> nmz/ mfn ~dhf of ao}xon F2&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
44/58
>>
Prhjmx~okfz
^doz z~hp cxz~ bh nkfh mz zkkf mz ~dh jhaaz mrh of grhh|ofi chnom& NCZK mfn
zkch k~dhr jrkprk~hj~mf~z mrh ~ktoj ~k jhaaz mfn zk zdkxan fk~ bh htpkzhn ~k
~dh jhaaz m~ rkkc ~hcphrm~xrh gkr mf akfihr ~dmf fhjhzzmr& ^dmyofi kg ~dh
womaz mfn pamjofi kg ~dh jhaa zxzphfzokf bmje of~k jxa~xrh chnom zdkxan mazk bh
nkfh whr }xojea gkr ~dh zmch rhmzkfz&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
45/58
>8
Htphrochf~ B9
RHWOWMA KG JHAA AOFHZ
MocCmf jxa~xrhz kb~mofhn grkc m jxa~xrh jkaahj~okf yoaa mrrowh grk|hf mfn of krnhr
~k xzh ~dhc! ~dh jhaaz cxz~ bh ~dmyhn mfn px~ of~k jxa~xrh& O~ oz wo~ma ~k ~dmy
jhaaz jkrrhj~a of krnhr ~k cmof~mof ~dh womboao~ kg ~dh jxa~xrh mfn hfmbah ~dh
jxa~xrh ~k rhjkwhr ckrh }xojea& Zkch jrkprk~hj~mf~z zxjd mz NCZK mrh ~ktoj
mbkwh > J& ^dhrhgkrh o~ oz hzzhf~oma ~dm~ ~dh jxa~xrhz bh ~dmyhn }xojea noax~hn
of jxa~xrh chnoxc ~k cofoco|h ~dh ~ktoj hgghj~z&
Cm~hromaz rh}xorhn
Chnom# prh#ymrchn ~k ~dh mpprkprom~h ~hcphrm~xrh&
H}xopchf~ rh}xorhn
Phrzkfma prk~hj~owh h}xopchf~ $z~hroah iakwhz! ambkrm~kr jkm~/
Ym~hrbm~d zh~ ~k mpprkprom~h ~hcphrm~xrh
Cojrkbokakiojma zmgh~ jmbofh~ m~ mpprkprom~h jkf~mofchf~ ahwha JK2 ofjxbm~kr
Prh ambhahn gamzez
Cmrehr phf
Cojrkpoph~~hz
Mcpxah rmje
^ozzxh
Prkjhnxrh
=& ^meh ~dh jhaaz grkc #40 J& ^dmy o~ m~ :9 J
2& Jhf~rogxih m~ 2000 rpc gkr 8 cof
:& Nozjmrn ~dh zxphrfm~mf~&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
46/58
>7
>& ^rmfzghr ~k =8 ca ~xbh& Mnn 8ca 20( NCHC
8& Jhf~rogxih m~ 2000 rpc gkr 8 cof mfn nozjmrn ~dh zxphrfm~mf~&
7& ^meh 8 ca 20 ( NCHC of jxa~xrh gamze& ^rmfzghr phaah~ ~k o~
9& Z~mr~ cmof~mofofi ~dh jhaa aofh
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
47/58
>9
Htphrochf~ B4
JHAA ZHHNOFI GKR NFM+RFM OZKAM^OKF
Prkjhnxrh
=& Nozjmrn htoz~ofi chnom
2& Mnn 2 ca PBZ#HN^M mfn ymzd ~yojh
:& Mnn ~rpzof mfn ihf~a zyora
>& Mnn = ca chnom
8&^rmfzghr ~k Jhf~rogxih ~xbh
7& Jhf~rogxih m~ =000rpc gkr : cof
9& Nozjmrn zxphrfm~mf~4& Mnn =ca grhzd chnom
?& Noax~h o~ =0 gkan yo~d chnom&
=0& ^meh =0xa mfn jkxf~ ~dh fxcbhr kg jhaaz xzofi dmhckj~kch~hr&
==& Yh rh}xorh : t =08 jhaaz of m jxa~xrh pam~h& Zk! yo~d ~dh dhap kg
W=F=3W2F2
Yh jmajxam~h ~dh wkaxch kg chnom ~k bh mnnhn ~k hmjd jxa~xrh pam~h&
=2& Mnn U=000#$wkaxch kg jhaa mnnhn/ S chnom ~k hmjd jxa~xrh pam~h
=:& Ofjxbm~h m~ :9J
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
48/58
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
49/58
>?
Htphrochf~ B=0
NFM OZKAM^OKF GRKC JXA^XRHN JHAA AOFH
$]OMIHFNFMozkam~okfeo~/
=& Jhaaz zxzphfnhn of NCHC z~krhn m~ #20 J
2& Rhckwh ~dh zxphrfm~mf~ jkcpah~ha mfn nozjmrn
:& Mnn 20xa PBZ ~k rhzxzphfn ~dh phaah~
>& Mnn 20xa prk~hofmzh E
8& Mnn 20xa bxgghr MA $azoz bxgghr/ mfn cot b pxazh wkr~htofi gkr =8z
7& Ofjxbm~h m~ 87 J gkr =0 cof
9& Mnn 200xa jdoaahn majkdka $70#=00(/ ' cot b pxazh wkr~htofi gkr =8z
4&^rmfzghr ~k cofo zpof jkaxcf ' jhf~rogxih m~ 4000rpc! 2:#28 J gkr 2
cof
?& Nozjmrn ~dh goa~rm~h
=0& Mnn Bxgghr MY=$ymzd bxgghr/ 800 xa ' jhf~rogxih m~ 4000rpc! 2:
28 J gkr 2 cofx~hz
==& Nozjmrn ~dh goa~rm~h
=2& Mnn 800xa Bxgghr MY2$ymzd bxgghr/
=:& Jhf~rogxih m~ =>000 rpc m~ 28 J gkr : cof
=>& Mnn 200xa Bxgghr MH$hax~okf bxgghr/
=8& Ofjxbm~h m~ rkkc ~hcphrm~xrh gkr 8 cof
=7& Jhf~rogxih m~ 4000 rpc m~ 28 J gkr 2 cof
=9& Z~krh m~ #20 J
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
50/58
80
Htphrochf~ B==
]XMF^OGOJM^OKF XZOFI FMFKNRKP
Moc
Pxro~ kg NFM zmcpah jmf mazk bh jmajxam~hn bmzhn xpkf o~z mbzkrbmfjh kg
aoid~& M pxrh zmcpah kg NFM dmz m 270fc+240fc rm~ok kg =&4& Rm~okz nhwom~ofi
grkc ~doz xzxmaa ofnojm~h jkf~mcofm~okf kg ~dh zmcpah yo~d prk~hofz! krimfoj
zkawhf~z kr RFM kr jkxan ofnojm~h nhirmnm~okf kg ~dh NFM zmcpah&
Cm~hromaz Rh}xorhn
=&Fmfknrkp
Prkjhnxrh
=& Jahmf ~dh zmcpah akmnofi pkof~ yo~d z~hroah ym~hr ~k ofo~omao|h ~dh
h}xopchf~
2& Kphf ~dh bxfnahn zkg~ymrh
:& Mnn =xa kg fxjahmzh grhh ym~hr zkax~okf of ydojd NFM oz nozzkawhn mfn
jmajxam~h Bamfe
>& Fky jahmf ~dh akmnofi pkof~ mfn akmn zxbzh}xhf~ zmcpahz mfn chmzxrh
}xmf~ogojm~okf&
8& Rhjkrn ~dh wmaxhz kg NFM jkfjhf~rm~okf$fi+xa/ mfn M270+M240
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
51/58
8=
Htphrochf~ B=2
RFM OZKAM^OKF GRKC JXA^XRHN JHAA AOFHZ $^RO\KA
CH^DKN/
Prkjhnxrh
=& Ymzd jhaaz yo~d =T PBZ&
2& Mnn =ca ^RO|ka phr yhaa&
:& Ofjxbm~h m~ rkkc ~hcphrm~xrh gkr : cofx~hz
>& Mnn 200xa JDJa:
8& Pxazh wkr~ht gkr =8 zhjkfnz mfn ofjxbm~h m~ rkkc ~hcphrm~xrh gkr :
cofx~hz&7& Jhf~rogxih gkr =8 cofx~hz m~ 4000 rpc m~ > J&
9& Poph~~h ym~hr pdmzh of~k m fhy hpphfnkrg ~xbh&
4& Mnn 800xa ozkprkpmfka phr ca kg ~ro|ka
?& Ofjxbm~h m~ #20 J gkr :0cofx~hz&
=0&Jhf~rogxih gkr =0 cofx~hz m~ =>000rpc m~ > J
==&Ymzd phaah~ yo~d 800xa 90( h~dmfka&
=2& Jhf~rogxih gkr 8 cof m~ =>000rpc m~ rkkc ~hcphrm~xrh&
=:& Mor nr ~dh phaah~ $20 cof/
=>& Rhzxzphfn of :0#80xa fxjahmzh#grhh ym~hr&
=8&Ehhp m~ 87 J gkr =0 cofx~hz
=7&Z~krh m~ #40J&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
52/58
82
Htphrochf~# B=:
MZZHZZCHF^ KG ]XMAO^_ MFN ]XMF^O^_ KG RFM&
m/ ]xmao~ kg RFM jmf bh jdhjehn b mimrkzh hahj~rkpdkrhzoz gkaakyhn b
h~donoxc brkconh z~mofofi& 2a kg RFM oz rxf kf =&2( mimrkzh iha mfn
pdk~kirmpdhn&
^dh prhzhfjh kg jrozp bmfnz jkrrhzpkfnofi ~k 24 Z rRFM mfn =4 Z rRFM
ofnojm~hz ~dh }xmao~ kg ~dh RFM ozkam~hn&
b/ RFM jkfjhf~rm~okf jmf bh chmzxrhn xzofi FmfkNrkp^C
zphj~rkpdk~kch~hr&
M270+M240 rm~ok gkr RFM zdkxan bh =&4#2& M270+M2:0 rm~ok zdkxan bh 2#2&2
Prkjhnxrh
=& Jahmf ~dh zmcpah akmnofi pkof~ yo~d z~hroah ym~hr&
2& Kphf ~dh bxfnahn zkg~ymrh
:& Mnn =xa kg fxjahmzh grhh ym~hr zkax~okf of ydojd RFM oz nozzkawhn mfn
jmajxam~h Bamfe
>& Fky jahmf ~dh akmnofi pkof~ mfn akmn zxbzh}xhf~ zmcpahz mfn chmzxrh
}xmf~ogojm~okf&
8& Rhjkrn ~dh wmaxhz kg RFM jkfjhf~rm~okf M270+M240 mfn M270+M2:0&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
53/58
8:
Htphrochf~ B=>
jNFM PRHPMRM^OKF
Prkjhnxrh
=& Prhpmrh m cot~xrh of mf hpphfnkrg ~xbh yo~d ~dh gkaakyofi jkfz~o~xhf~z
=0 t R^ bxgghr 2 xa
28T nF^P 0&4 xa
=0 T rmfnkc prochr 2 xa
Rhwhrzh ~rmfzjrop~mzh = xa
Fxjahmzh Grhh Ym~hr >&2 xa
=0xa
2& Prhpmrh = xi kg RFM of =0 xa
:& Cot =0xa kg RFM mfn =0 xa kg rhmj~okf cot&
>& Pamjh ~dh ~xbh of m ~dhrcma jjahr mfn rxf ~dh gkaakyofi prkirmc
28 kJ =0 cof
:9 kJ =20 cof
48 kJ 8 cof
> kJ ofgofo~
]XMAO^_ JKF^RKA ^HZ^ GKR jNFM
^doz ~hz~ ymz bhofi phrgkrchn b PJR mcpaogojm~okf kg#Mj~of&
Rhmj~okf cot~xrh ymz prhpmrhn yo~d gkaakyofi jkcpkfhf~z
jNFM # =A
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
54/58
8>
=0TR^ bxgghr # =A P&^&K
2&8cCnF^P # =A
20 xC gkrymrn prochr # 0&0?xa
20 xC rhwhrzh prochr # 0&0?xa
^m} pkachrmzh # 0&2xa
D2K # 7&72xa
=0xa
PJR jkfno~okfz
?8`J : cof
?8J :0 zhj
87&9J =8 zhj
92J :0 zhj
92J =0 cof
>J
Gkaakyofi PJR!~dh prknxj~ ymz rxf of =( iha mfn mfmazhn gkr bmfnz
jkrrhzpkfnofi ~k B#mj~of&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
55/58
88
Htphrochf~ B=8RHMA#^OCH PJR
Moc
^doz ~hjdfo}xh oz xzhn ~k mcpaog mfn zocxa~mfhkxza }xmf~og m
~mrih~hn NFM ckahjxah& Dhrh ~dh mcpaogohn NFM oz nh~hj~hn mz ~dh rhmj~okf
prkirhzzhz of rhma ~och&^yk jkcckf ch~dknz gkr nh~hj~okf kg prknxj~z of rhma#
~och PJR mrh< $=/ fkf#zphjogoj gaxkrhzjhf~ nhz ~dm~ of~hrjmam~h yo~d mf
nkxbah#z~rmfnhn NFM! mfn $2/ zh}xhfjh#zphjogoj NFM prkbhz jkfzoz~ofikg kaoikfxjahk~onhz ~dm~ mrh ambhahn yo~d m gaxkrhzjhf~ rhpkr~hr ydojd phrco~z
nh~hj~okf kfa mg~hr dbrono|m~okf kg ~dh prkbh yo~d o~z jkcpahchf~mr NFM
~mrih~&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
56/58
87
^dhrh mrh bmzojmaa ~yk ch~dknz kg mfma|ofi ~dh nm~m grkc m rhma ~och PJR#
mbzkax~h }xmf~ogojm~okf mfn rham~owh }xmf~ogojm~okf& Rham~owh }xmf~ogojm~okf kr
jkcpmrm~owh }xmf~ogojm~okf chmzxrhz ~dh rham~owh jdmfih of cRFM htprhzzokf
ahwhaz& O~ nh~hrcofhz ~dh jdmfihz of z~hmn z~m~h cRFM ahwhaz kg m ihfh mjrkzz
cxa~opah zmcpahz mfn htprhzzhz o~ rham~owh ~k ~dh ahwhaz kg mfk~dhr RFM& Rham~owh
jkfjhf~rm~okfz kg NFM prhzhf~ nxrofi ~dh htpkfhf~oma pdmzh kg ~dh rhmj~okf mrh
nh~hrcofhn b pak~~ofi gaxkrhzjhfjh mimofz~ jjah fxcbhr kf m akimro~dcoj
zjmah $zk ~dm~ mf htpkfhf~omaa#ofjrhmzofi }xmf~o~ yoaa zdky mz m z~rmoid~ aofh/&
M ~drhzdkan gkr nh~hj~okf kg gaxkrhzjhfjh mbkwh bmjeirkxfn oz nh~hrcofhn& ^dh
jjah m~ ydojd ~dh gaxkrhzjhfjh grkc m zmcpah jrkzzhz ~dh ~drhzdkan oz jmaahn
~dh jjah ~drhzdkan! J~&
^dh mckxf~ kg ~mrih~! fkrcmao|hn ~k mf hfnkihfkxz rhghrhfjh mfn rham~owh ~k m
jmaobrm~kr oz iowhf b!
Mckxf~ kg ~mrih~ 3 2#J^
Ydhrh J^3 $J^!~mrih~ J^!mj~of /~och!t $J^!~mrih~ J^!mj~of/~och!0&
Dhrh ~och t oz mf ~och pkof~ mfn ~och0 rhprhzhf~z ~dh htprhzzokf kg ~dh
~mrih~ ihfh fkrcmao|hn ~k #mj~of&
Prkjhnxrh
PJR jkcpkfhf~z gkr ^m}Cmf bmzhn ihfh htprhzzokf mzzm
jNFM # 2a
^m}Cmf Xfowhrzma PJR Cmz~hr Cot # 8 a
20T ^m}Cmf Ihfh Htprhzzokf Mzzm Cot # 0&8 a
Fxjahmzh#grhh ym~hr # 2&8 a
^k~ma # =0 a
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
57/58
89
Rhmj~okf cot ymz prhpmrhn gkr B#mj~of mazk! ydojd zhrwhn mz ~dh rhghrhfjh
ihfh&
^dh zmcpahz yhrh akmnhn kf ?7#yhaa pam~h mfn hmjd zmcpah ymz rxf of
~ropaojm~hz&
^dh pam~h ymz rxf kf ~dh MBO 9?00D^ yo~d ~dh gkaakyofi zh~~ofiz0 jjahz kg
Z~hp :&=8 zhj m~ ?8 J mfn ~dhf = cofx~h m~ 70 J&
^dh rhzxa~z yhrh mfma|hn xzofi ZNZ R] Cmfmihr zkg~ymrh&
&
7/31/2019 SNP Genotyping and Animal Cell Culture techniques
58/58
Rhghrhfjh