Molecular Phylogenetic relationships among the species of geckos in Adams’peak in Sri Lanka
•Download as PPT, PDF•
0 likes•256 views
A ten month survey was undertaken to document the diversity and abundance of lizards at the Adam’s peak in Sri Lanka. We use 700 bp of mitochondrial (cytb) and nuclear (RAG1 and Phosducine) DNA sequence data to recover phylogenetic relationships and our well resolved tree, to elucidate a biographic pattern and close relationship among lizards. Our data demonstrate the phylogenetic utility of Phosducine, a noval marker in squmate phylogenetics at the intrageneric level.
Recommended
Recommended
More Related Content
Viewers also liked
Viewers also liked (12)
Similar to Molecular Phylogenetic relationships among the species of geckos in Adams’peak in Sri Lanka
Similar to Molecular Phylogenetic relationships among the species of geckos in Adams’peak in Sri Lanka (20)
Recently uploaded
Recently uploaded (20)
Molecular Phylogenetic relationships among the species of geckos in Adams’peak in Sri Lanka
- 2. Green Anole Anolis carolinensis Brown Anole (exotic) Anolis sagrei sagrei Southeastern Five-lined Skink Eumeces inexpectatus Indo-Pacific Gecko (exotic) Hemidactylus garnoti Red-headed rock agama, Agama agama Six-lined Racerunner Cnemidophorus sexlineatus
- 3. Determination of the Diversity Of Lizard Species by using Mitochondrial DNA
- 4. •Extraction of DNA from Lizard’s Tails •Polymerase Chain Reaction (PCR) •Analysis of PCR Products
- 5. Cleaned Lizard Tail Sample 100 ul Protinase Buffer + Protinase K Enzyme Overnight Incubation at 50 C Digested Sample 3 min Centrifuge at 12000 rpm For Supernatant RNAse for 15 minIncubation at 37 C Digested Sample Iso - Propanol Centrifuge at 13000 rpm For 3 min Pellette (Discard the supernatant) 70 % ETOH Centrifuge at 3 min 13000 rpm For Pellette (Discard the supernatant) Incubation at 56 C TE Buffer Extracted DNA
- 6. Master Mix Preparation Distilled Water 14 ul 10x Buffer 2.5 ul 2.5mM dNTP’s 2.5 ul 5uM Primer 2.5 ul Taq DNA Polymerase 0.5 ul Template DNA 3 ul PCR Protocol 95 C 3 min 95 C 30 sec 50 C 30 sec 72 C 1 min 72 C 3 min 35 Cycles PRIMERPRIMER GENEGENE SEQUENCESEQUENCE CytbF700CytbF700 Cytochrome bCytochrome b 5’CTTCCAACACCAYCAAACATCTCAGCATG5’CTTCCAACACCAYCAAACATCTCAGCATG ATGAAA 3’ATGAAA 3’ CytbR700CytbR700 Cytochrome bCytochrome b 5’ACTGTAGCCCCTCAGAATGATATTTGTC5’ACTGTAGCCCCTCAGAATGATATTTGTC CTCA 3’CTCA 3’ PHOF2PHOF2 PhosducinPhosducin 5’ AGATGAGCATGCAGGAGTATGA 3’5’ AGATGAGCATGCAGGAGTATGA 3’ PHOR1PHOR1 PhosducinPhosducin 5’ TCCACATCCACAGCAAAAAACTCCT 3’5’ TCCACATCCACAGCAAAAAACTCCT 3’ RAG1F700RAG1F700 RAG-1RAG-1 5’ GGAGACATGGACACAATCCATCCTAC 3’5’ GGAGACATGGACACAATCCATCCTAC 3’ RAG1R700RAG1R700 RAG-1RAG-1 5’ TTTGTACTGAGATGGATCTTTTTGCA 3’5’ TTTGTACTGAGATGGATCTTTTTGCA 3’
- 8. 100 bp ladder AA 00 AA 01 AA 02 AA 04 AA 05 ID NoID No BAND SIZEBAND SIZE RESULTSRESULTS DATEDATE AA 00AA 00 373bp373bp PositivePositive 20. 02. 200820. 02. 2008 AA 01AA 01 373bp373bp PositivePositive 20. 02. 200820. 02. 2008 AA 02AA 02 373bp373bp PositivePositive 20. 02. 200820. 02. 2008 AA 04AA 04 373bp373bp PositivePositive 20. 02. 200820. 02. 2008 AA 05AA 05 373bp373bp PositivePositive 20. 02. 200820. 02. 2008
- 9. AGAROSE GEL ELECTROPHORESIS SEQUENCING Required Band size (373 bp) was observed for recently analyzed 16 samples. Characterization of Lizards by there Mitochondrial DNA