aflp1.ppt

7/26/2019 aflp1.ppt

1/55

9/22/2012 Mithilesh Singh, M Sc.(GPB) 1

welcomewelcome

7/26/2019 aflp1.ppt

2/55

Mithilesh KumarMithilesh Kumar

SinghSingh Sr. M sc.(GPB) Sr. M sc.(GPB)

PALB 1205 PALB 12059/22/2012 2Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

3/55

What is marker ?

haracters !nheritance "#ll#$e%

&i'erent classes #"markers

M#rh#l#gical Bi#chemical markers

9/22/2012

3

Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

4/55

Markers at !" le#elMarkers at !" le#el

came to the resc$ecame to the resc$e

9/22/20124 Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

5/55

AbundantAbundant

Highly polymorphicHighly polymorphic

No environmental influenceNo environmental influence

No influence of ontogeny of individualNo influence of ontogeny of individual

Co-dominant or dominantCo-dominant or dominant


7/26/2019 aflp1.ppt

6/55

%hanging relati#e

im&ortance o' i''erent

molec$lar markers

!at$re e#iews Genetics

*ol$me + 200- Page 9/22/20126 Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

7/55

7/26/2019 aflp1.ppt

8/55

Single loc$s

M$lti loci

9/22/2012* Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

9/55

#arer! %&'"

#ini!atellit

e! %A"D $$% ($$% $$C" CA"$ $CA% A&'"

Abundance

'evel ofpolymorphi!m

'ocu! !pecificity

Codominance ofallele!

%eproducibility

'abour-inten!ity

+echnicaldemand!

,perational co!t!

Developmentco!t!

uantity of DNAre.uired

Amenability toautomation

igh Mei$m ow 3es !o

Compari!on of DNA marer! for their efficiency

9/22/2012/ Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

10/55

What is AFLP ?

"m&li'ie 'ragment length &ol4mor&hism

("P) are generate 54 com&lete("P) are generate 54 com&lete

restriction enon$clease igestion o'restriction enon$clease igestion o'

genomic !" 'ollowe 54 selecti#egenomic !" 'ollowe 54 selecti#e

ream&li'ication an electro&horesis o' aream&li'ication an electro&horesis o' a

s$5set o' 'ragments an res$lting ins$5set o' 'ragments an res$lting in

$ni6$e, re&ro$ci5le 'inger&rint 'or each$ni6$e, re&ro$ci5le 'inger&rint 'or eachini#i$alini#i$al..

9/22/2012Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

11/55

Contd.

P *ase% ( +#s et al., 1--5)

LP / P

#*ust

aster, less la*#ur intensie

m#re in"#rmati#n. &As #" an #rigin #r

c#mle3it.


00

7/26/2019 aflp1.ppt

12/55

Stes in ALP4

Dige!tion

Adaptor 'igation

Amplification

1lectrophore!i!

9/22/2012

02


7/26/2019 aflp1.ppt

13/55

03

%e!triction fragment%e!triction fragment

enomic DNAenomic DNA

%are cutter EcoRI &re.uent cutter Mse I

$tep ( %e!triction Dige!tion

confirmation


7/26/2019 aflp1.ppt

14/55

(iii)la5el one stran o' the s P% &ro$cts,

&re#ents the occ$rrence o' 7o$5lets7 on the gels8ne6$al mo5ilit4 o' the two strans o' the am&li'ie 'ragments.

(ii) 'ragments to 5e am&li'ie is re$ce.

(i) he 're6$ent c$tter generate small !" 'ragments.

%ationale

9/22/2012 1Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

15/55

1+

estriction ragmentAdapter(ECORI)Adapter

Mse i)

%e!triction fragment! 7ith adapter! are called

a! +agged %e!triction &ragment! +%&

$tep (( 'igation of Adapter!$tep (( 'igation of Adapter!


7/26/2019 aflp1.ppt

16/55

1

%e!triction &ragmentAdapter "a&ter

estriction ragment "a&ter"a&ter

%e!triction &ragmentAdapter Adapter

+ype! of +%&!+ype! of +%&!

confirmation


7/26/2019 aflp1.ppt

17/55

Contd.

$# %i'erent a%at#rs.

target sites "#r rimer

annealing

c#re se6uence and en7me8seci9c se6uence

9/22/2012

0)


7/26/2019 aflp1.ppt

18/55

1co%(-adapter

+-%%G"G"%G%G"%%-

-%"%G"%G%"GG""-+

+-G"%G"G"G%%G"G-

-"%%"GG"%%"-+

#!e(-adapter

9/22/2012 1:Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

19/55

estricti#n8ligati#n

9/22/2012

0/


7/26/2019 aflp1.ppt

20/55

1co%( "%1-$1'1C+(81 "%(#1%

#!e( "%1-$1'1C+(81 "%(#1%

G"G"%G%G"%% "" %"

%" " G"G%%G"G"

9/22/2012 20Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

21/55

S;;%

7/26/2019 aflp1.ppt

22/55

Mse>>>>

)i@!ult to s!ore

9/22/2012

40


7/26/2019 aflp1.ppt

42/55

'inger&rint &atterns are #er4 similar

$sing i''erent tem&late 6$antities

(2+ ng to 2+ &g).

Insensitivit to template!on!entration


7/26/2019 aflp1.ppt

43/55

a5ele &rimer $se limite 6$antit4Primer is e@ha$ste."m&li'ication sto&s.'$rther thermo c4cling oes not a''ect the 5an &attern

his characteristic is elegantl4 $tilie in the "P

&rotocol, which $ses an e@cess o' P% c4cles.

:ual intensit from varia#letemplate !on!entration


7/26/2019 aflp1.ppt

44/55

hy #!e( hy not +a.(

;$kar4otic !"s "-richMsel ""a6< %G"

hy 1co%(

%eliable lo7 co!t$i?-cutter enymeComplete re!triction of the DNA


7/26/2019 aflp1.ppt

45/55

9/22/2012 Mithilesh Singh, M Sc.(GPB) +

Application of A&'"

in crop improvement

7/26/2019 aflp1.ppt

46/55


'inage mapping

"arentage analy!i!

#ea!uring genetic diver!ity

"opulation genetic!

Development of !ingle locu! marer!

from A&'" for #A$

A&'" application!A&'" application!

7/26/2019 aflp1.ppt

47/55

C

Ca!e !tudy

%elation!hip bet7een hybridperformance and A&'" ba!ed genetic

di!tance in highland maie inbred

line!E

'ege!!e et al., 2==)

http://www.springer.com/life+sci/plant+sciences?SGWID=0-10038-0-0-0

7/26/2019 aflp1.ppt

48/55

:9/22/2012 Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

49/55


7/26/2019 aflp1.ppt

50/55

+09/22/2012 Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

51/55

+19/22/2012 Mithilesh Singh, M Sc.(GPB)

Dendrogram derived from >"#A analy!i! ba!ed on 32 maie

7/26/2019 aflp1.ppt

52/55

+2

Dendrogram derived from >"#A analy!i! ba!ed on 32 maie

inbred line!


7/26/2019 aflp1.ppt

53/55

+

%oncl$sion

Signi'icant &ositi#e correlation mani'este

5etween Gs an h45ri &er'ormance 'or most o'

the traits in in5re line @ tester com5inations9/22/2012 Mithilesh Singh, M Sc.(GPB)

7/26/2019 aflp1.ppt

54/55

54

Conclu!ion

&ar from being almo!t forgottenE A&'" i! a

highly u!eful techni.ue and if fo!tered byparallel development of ne7 analy!i!

method! 7ill continue to be at the forefront

in an!7ering important !cientific .ue!tion!

!


Bou

7/26/2019 aflp1.ppt

55/55

han$ing

Bou

Documents

aflp1.ppt