Molecular

4. RAPDRAPD::
11Genomic DNAGenomic DNA
22PCRPCRprimersprimers
33
44patternspatternsprimersprimers
55patternspatterns
Randam Amplified Polymorphic DNARandam Amplified Polymorphic DNA ((RAPDRAPD:(:(
66

5. 11Genomic DNAGenomic DNA

6. 22PCRPCRprimersprimers
MgClMgCl22
Template DNATemplate DNA..
Two oligonucleotide primersTwo oligonucleotide primers..
Taq polymeraseTaq polymerase..
dNTPsdNTPs..
PCRPCR::

8. one primerone primer
1010
DNADNA

9. --RAPDRAPD331010
DNADNA..
AgaroseAgarose
ethidium bromideethidium bromide..
33

10. 200–2000bp
2annealing sites
PrimerPrimerRAPDRAPD
22annealing sitesannealing sites
primerprimer
200200––20002000bpbp..

11. BandsBandsDNADNAprimerprimer
√
L3
X L4 L5
L2
L6
L1
X √
A B
primerprimerDNADNA..
PCR Reaction
BA
PCR Product
11,,4422annealing sitesannealing sitesPCRPCR
22,,44primerprimerextensionextension
33,,55primerprimerextensionextension
A bandA band22,,55primerprimer.. B bandB band33,,66primerprimer..

12. LL66LL44 LL55
LL33LL11
XX XX XX √√
BB
PCR Reaction
PCR Product
BB

13. MM P 2P 2
200 bp200 bp
400 bp400 bp
100 bp100 bp
P 1P 1
500 bp500 bp
600 bp600 bp
700 bp700 bp
800 bp800 bp
300 bp300 bp
--
++ Agarose gelAgarose gel

14. 44patternspatternsprimersprimers
11 3322--
++
11 3322--
++
11 3322--
++
MonomorphicMonomorphic MonomorphicMonomorphic PolymorphicPolymorphic
))))No FragmentNo Fragment

15. 55patternspatterns
200bp.
300bp.

16. 66
1 = Presence1 = Presence
markermarkerbandsbands
0 = Absence0 = Absence
--Similarity matrixSimilarity matrix..
--Cluster analysisCluster analysis ((DendrogramDendrogram(.(.

17. V.2 V.2.S.1 V.2.S.2 V.2.S.3 V.2.S.4 V.3 V.3.S.1 V.3.S.2 V.4 V.4.S.1 V.4.S.2 V.4.S.3 V.4.S.4
V.2 1
V.2.S.1 0.52 1
V.2.S.2 0.48 0.63 1
V.2.S.3 0.56 0.48 0.55 1
V.2.S.4 0.45 0.35 0.39 0.46 1
V.3 0.51 0.49 0.51 0.6 0.64 1
V.3.S.1 0.56 0.52 0.52 0.57 0.53 0.72 1
V.3.S.2 0.59 0.50 0.56 0.62 0.55 0.69 0.66 1
V.4 0.50 0.44 0.45 0.53 0.61 0.70 0.63 0.66 1
V.4.S.1 0.33 0.51 0.51 0.42 0.27 0.35 0.36 0.41 0.38 1
V.4.S.2 0.45 0.52 0.60 0.58 0.37 0.53 0.50 0.57 0.53 0.62 1
V.4.S.3 0.44 0.55 0.55 0.51 0.36 0.48 0.47 0.54 0.47 0.58 0.71 1
V.4.S.4 0.39 0.53 0.53 0.47 0.30 0.42 0.42 0.46 0.41 0.53 0.56 0.59 1
--Similarity matrixSimilarity matrix::

18. 024681012141618
0.24
0.32
0.4
0.48
0.56
0.64
0.72
0.8
0.88
0.96
Similarity
V.1.S.2
V.1.S.3
V.2.S.1
V.2.S.2
V.4.S.1
V.4.S.2
V.4.S.3
V.4.S.4
V.2.S.3
V.2
V.2.S.4
V.1
Cham
Veery
V.3
V.3.S.1
V.3.S.2
V.4
V.1.S.1
--Cluster analysisCluster analysis ((DendrogramDendrogram(:(:

19. Why choosing RAPD techniqueWhy choosing RAPD technique??
•RAPDRAPD
probeprobe
•

20. •
--Taxonomic studiesTaxonomic studies..
--Genome mappingGenome mapping..
--DNA finger printingDNA finger printing..
--Varietal identificationVarietal identification..
--Gene tagging (QTLs mappingGene tagging (QTLs mapping((..
--Somaclonal identificationSomaclonal identification..
--Germplasm diversityGermplasm diversity..
markermarker

21. RAPDRAPD
11markermarker
•
•
•QTL analysisQTL analysis.(.(

22. Genotype identificationGenotype identification..
22
Relatedness studiesRelatedness studies..
Individual genotypesIndividual genotypes..

23. RAPDRAPD::
11
22
DNADNA
33
44
55DNADNA..

24. RAPDRAPD::
11
22DNADNA
33DNADNA..
44markermarker