| Tutorial |
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DNA can be extracted from whole blood using a standard phenol/chloroform extraction method. A battery of 25 microsatellite markers which are based on the guidelines of ISAG & FAO’s DAD-IS program can be utilised to generate allelic data. Only forward primers at 5' end of each pair needs labelling with one of the four fluorophores, i.e., FAM (blue), VIC (green), NED (yellow) and PET (red). For details please see Kumar et al., 2009.
PCR reactions may be carried out using
50–100 ng genomic DNA in a 25 µl reaction volume.
A touchdown PCR protocol can be used
with initial denaturation of 95 ◦ C for 1 min, 3 cycles of 95 ◦ C for 45 s, and
60 ◦ C for 1 min, 3 cycles of 95 ◦ C for 45 s, and 57 ◦ C for 1 min, 3 cycles of
95 ◦ C for 45 s, and 54 ◦ C for 1 min, 3 cycles of 95 ◦ C for 45 s and 51 ◦ C
for 1 min, 20 cycles of 95 ◦ C for 45 s, and 48 ◦ C for 1 min with final
extension at 72 ◦ C for 5 min. At the end
reaction can be stopped by adding 5.0 µl of stop dye (95% formamide, 0.25% bromophenol blue
and 0.25% xylene cyanol). 6 µl of PCR
products can be loaded on a 2% agarose gel, for electrophoresis to visualize it
in UV light after ethidium bromide staining to detect the amplification. The PCR products can be mixed in a ratio of 1:1.5:2:2 of FAM, VIC, NED and PET labeled primers, respectively. 0.5 µl of this mixture (a post PCR multiplex of 4–5 primers) can be combined with 0.3 µl of Liz 500 as internal lane standard of Applied Biosystems, U.S.A. and 9.20 µl of Hi-Di Formamide per sample. The resulting mixture should be denatured by incubation for 5 min at 95 ◦ C. These denatured samples have to be run on automated DNA sequencer of Applied Biosystems (ABI 3100 Avant). The electropherograms drawn through Gene Scan can be used to extract DNA fragment sizing details using Gene Mapper software (version 3.0) (Applied Biosystems, U.S.A.). |
Schematic Diagram |
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How to use BIS - Goat Web Server? |
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| The purpose of BIS - Goat server is to predict the Goat breed based on input data. The Homepage has seven different tabs viz. Home, Submission, Algorithm, Tutorial, Team, Links, FAQs. To use the BIS - Goat server to predict the goat breed, Submission tab needs to be used. Algorithm tab gives the details of the algorithm used for the development of BIS - Goat web-server. Help page is also available for assisting the users. The diagrammatical Illustration of how to use this server is as follows: | |
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| Input Methods : Data can be input either by manually entering the data in the form or by uploading a .txt or .csv file with 18 values. | |
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| You can use sample data for first view of the prediction result. Just click on use sample data button it will automatically fill your input form. | |
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| after that click on Submit Query button for the prediction result. | |
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| The result gives the probability of a particular breed or breeds, goat image from data provided.To know more about the prediction result please click on goat image. | |
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| Scroll down will give you more about the goat. | |
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