1. MOLECULAR GENETIC CHARACTERIZATION OF DAHLEM RED LAYERS MOLECULAR GENETIC CHARACTERIZATION OF DAHLEM RED LAYERS Ch.Shivaprasad Assistant Professor (AGB) ILFC; C.V.Sc. Korutla,SVVU, Telangana, India

2. INTRODUCTION It is a layer type Big size brown colored egg Disease resistant Back yard poultry production DAHLEM RED BIRDS

3. OBJECTIVES  Molecular genetic characterization Dahlem Red layers using RAPD markers  To study the genetic similarity and genetic distance between full sib, half sib mated non-inbred groups

4. MATERIALS AND METHODS  A total of 48 birds  A total of 100 decamer primers serially numbered PDP40F to PDP115F and PDP155F to PDP 180F  Genomic DNA isolated from blood samples  Checking quality of DNA  Checking quantity of genomic DNA

5. RAPD-PCR ANALYSIS Amplification were carried out final volume of 12.5 μl 25 pM primer 200 μM of the dNTPs 2 mM Magnesium chloride 1.25 U Taq DNA polymerase 1.25 μl 10X PCR buffer 100 ng genomic DNA

6. PCR REACTION StepTemperatureDuration Initial denaturation95 o C2 minutes Cyclic denaturation94 o C1 minutes Primer annealing36 o C1 minutes 40 Cycles Primer extension72 o C2 minutes Final extension72 o C10 minutes Held at4 o CForever

7. Electrophoresis Visualized by UV transilluminator Gel documentation Analysis of Results by using data base software

8. STATISTICAL ANALYSIS The scoring and matching of bands-binary character matrix Band sharing- Nei’s method Band frequency- Raman et al. Genetics distance- Kuhnlein et al.

9. RESULTS Primer name Number of bands amplifiedSize range (bp) TotalPolymorphicMinimumMaximum PDP40F14103911592 PDP42F1243211660 PDP50F17122151788 PDP54F1992973223 PDP62F1473191530 PDP63F19112593408 PDP71F23181403158 PDP72F18132652872 PDP73F1792522480 PDP75F1483441711 PDP77F1552633409 PDP78F17101001542 PDP79F19101822054 PDP94F17122821752 PDP96F1491112176 PDP103F20182232797 PDP110F1251431966 PDP112F1572191697 PDP157F16102131866 PDPF168F1274502921 PDP176F17102621538 Total341204--

10. RAPD-PCR FINGERPRINTS Full sib-unique presence of 1866bp

11. Half sib-unique presence of 1788bp

12. Non-inbred-unique presence of 421pb

13. Intra-population genetic similarity based on band sharing Primer nameFull sibHalf sibNon-inbred PDP40F85.5085.8386.16 PDP42F92.5094.0090.66 PDP50F86.5088.1688.02 PDP54F88.0089.0089.66 PDP62F88.3388.5088.00 PDP63F85.3390.1694.50 PDP71F85.3386.6667.83 PDP72F88.8382.5080.83 PDP73F87.6691.0085.50 PDP75F85.6695.0096.00 PDP77F93.6689.3392.66 PDP78F89.0083.1680.16 PDP79F85.6688.8380.66 PDP94F94.3393.0075.16 PDP96F82.8382.0078.83 PDP103F74.1671.1675.83 PDP110F93.3387.5088.16 PDP112F79.6685.5077.33 PDP157F91.3391.6688.33 PDP168F97.0090.6677.50 PDP176F91.33 81.00 Mean87.9087.8583.94 SE1.14 1.58

14. Inter-population genetic similarities based on band sharing using pooled DNA Primer name Full sib and Half sib groups Half sib and Non- inbred groups Full sib and Non-inbred groups PDP 40F70.0080.0076.19 PDP 42F91.6684.61 PDP 50F85.7181.8186.95 PDP 54F88.2380.0092.5 PDP 62F96.2988.8876.92 PDP 63F94.1188.8889.47 PDP 71F73.6890.0073.68 PDP 72F89.6592.8596.77 PDP 73F93.3394.1187.50 PDP 75F91.8997.4389.47 PDP 77F87.5084.2193.33 PDP 78F100.0088.88 PDP 79F95.6572.7286.95 PDP 94F84.6181.4888.88 PDP 96F93.7591.4284.84 PDP 103F84.6192.8591.66 PDP 110F90.9095.65 PDP 112F94.7390.0094.73 PDP 157F100.0090.90 PDP 168F95.2395.6590.90 PDP 176F92.3096.5588.88 Mean90.1888.5188.07 SE1.641.431.34

15. Intra-population genetic similarities based on band frequencies Primer nameFull sibHalf sib Non-inbred PDP 40F75.0077.0881.81 PDP 42F89.5893.7597.72 PDP 50F85.0085.9387.50 PDP 54F83.8289.7085.52 PDP 62F80.3585.7183.33 PDP 63F80.3591.6698.33 PDP 71F95.0090.6267.04 PDP 72F85.2982.6971.15 PDP 73F82.1485.7185.29 PDP 75F88.4691.66100.00 PDP 77F92.8582.1490.00 PDP 78F78.8475.00 PDP 79F82.3580.8876.38 PDP 94F90.9090.3868.75 PDP 96F84.0981.2571.15 PDP 103F72.5070.8362.50 PDP 110F90.9084.0995.45 PDP 112F79.5480.4671.66 PDP 157F85.0088.4687.50 PDP 168F100.0092.5072.91 PDP 176F88.3386.5375.00 Mean85.2585.0981.14 SE1.451.332.43

16. Inter-population genetic similarities between groups based on band frequency Primer name Full sib and Half sib groups Half sib and Non-inbred groups Full sib Non-inbred groups PDP 40F60.2064.5659.45 PDP 42F99.3396.3397.00 PDP 50F82.5073.9089.25 PDP 54F87.0079.1083.39 PDP 62F88.7887.7383.16 PDP 63F81.6471.7765.68 PDP 71F72.6166.7267.31 PDP 72F68.6465.6257.88 PDP 73F79.4673.2478.42 PDP 75F83.0778.0091.42 PDP 77F82.6685.8089.26 PDP 78F76.4390.6072.70 PDP 79F90.3070.8975.54 PDP 94F79.9262.2957.05 PDP 96F75.5688.1869.02 PDP 103F51.1231.3319.60 PDP 110F80.3882.3587.35 PDP 112F80.3370.5569.06 PDP 157F76.6676.2868.68 PDP 168F90.5477.3377.91 PDP 176F83.2868.8869.35 Mean79.5374.3472.78 SE2.292.953.65

17. GENETIC DISTANCE GroupsFull sibHalf sibNon-inbred Full sib 0.87890.22900.3176 Half sib 0.87850.2963 Non-inbred 0.8394

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