840:153g:Projects/project24/2012/11/08

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(Autocreate 2012/11/08 Entry for 840:153g:Projects/project24)
Current revision (16:44, 29 November 2012) (view source)
(PCR and Gel electrophorosis fro extracted DNA sample)
 
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[[category:OWWLabNotebookV1]]
[[category:OWWLabNotebookV1]]
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On Tuesday, we ran DNA 3 sample at gel again because in last Agarose gel electrophorosis we didn't get band for DNA 3 however, the same sample had good ECORI digested band. We found that DNA 3 had similar intensity as DNA 2 and 5 so we mixed all three samples. We also,did PCR for our extracted DNA sample. This time we used non extended primers and run the samples at three different temperatures ( 65,55 and 50 degree centigrade). We also, tested for other group's DNA sample. To check if our primer is specific to our gene of interest or  not.
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On Tuesday, We did PCR for DNA samples with the extended primers to check if our extended primers is compatible with our new DNA sample at 3 temperature range 65ºC, 55ºC and 50ºC http://openwetware.org/wiki/Image:PIC_9.jpg. We also did agarose gel electrophoresis for PCR products. We got good results this time as well. Our DNA got amplified with extended primers and the band size was between 1300-1400bp which is the expected size of our gene of interest i.e. 1350bp. We also noticed that all the bands are same at different temperatures. We cut the band from gel and store them at -20ºC.  
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On Thursday, we did Agarose gel electrophorosis for our PCR product. This time we used 3ul marker. The result seems to be satisfactory this time because, we saw that our DNA was amplified with non- extended primers and also, it was specific to our DNA because there was no amplification on DNA of other groups. When we compared the band size with 100bp Marker, we found that the size of DNA band was around 1350bp which is the size of our gene of interest. So, we decided to do PCR of our 3 samples using extended primers in our next lab.
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On Thursday, We extracted DNA from agarose gel by using gel extraction kit and then run them in agarose gel.On agarose We saw that both our DNA6 and DNA mix gave good band of expected size i.e. 1350bp http://openwetware.org/wiki/Image:PIC_10.jpg. So in our next lab we will grow the plasmid culture.

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PCR and Gel electrophorosis fro extracted DNA sample

On Tuesday, We did PCR for DNA samples with the extended primers to check if our extended primers is compatible with our new DNA sample at 3 temperature range 65ºC, 55ºC and 50ºC http://openwetware.org/wiki/Image:PIC_9.jpg. We also did agarose gel electrophoresis for PCR products. We got good results this time as well. Our DNA got amplified with extended primers and the band size was between 1300-1400bp which is the expected size of our gene of interest i.e. 1350bp. We also noticed that all the bands are same at different temperatures. We cut the band from gel and store them at -20ºC.

On Thursday, We extracted DNA from agarose gel by using gel extraction kit and then run them in agarose gel.On agarose We saw that both our DNA6 and DNA mix gave good band of expected size i.e. 1350bp http://openwetware.org/wiki/Image:PIC_10.jpg. So in our next lab we will grow the plasmid culture.

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