Team:UNITN-Trento/Notebook/Labposts/06/25

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(Undo revision 78342 by Ggirelli (talk))
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{
{
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"date" : "2013-08-12",
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"date" : "2013-06-07",
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"author" : "fabio",
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"author" : "thomas-emil"
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"title" : "Blue light induction",
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"title" : "Second SAMsynthase extraction attempt - TOO SAD",
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"content" : "last week I was out for vacation, now I’ve just come back to the laboratory ready to start the blue light induction esperiment: Bruno gave me a blue light sensing device that produces blue pigment if induced. The part is composed by j23100-Yf1-FixJ-FixK2-CI-Plambda-amilCP. I need to make an inoculum o/n in order to proceed tomorrow.",
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"content" : "Since we didn't obtained our product using the Phusion PCR, we tried again to amplify the SAMsynthase gene from an extract of E.coli genomic DNA (strain MG1655). In order to do that we exploited the same primers used in our previous attempt (see 06-06-2013). For the protocol used see the <html><a href="https://2013.igem.org/Team:UNITN-Trento/Protocols#RBC-Taq-DNA-pol-PCR">RBC Taq PCR protocol</a></html>.. When the reaction finished, we tested the presence of the aplificate product througt an electrophoresis analisys (adding 2 µl of LD for 10 µl of DNA). <B>Results:</B> {{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel Image|<html> <img src="https://static.igem.org/mediawiki/2013/d/df/Tn-20130607-SAMsynth_taq.jpg" /> </html>}} As you can see from our gel image, our product is present in both reactions (TAQ and TAQ+Phusion). After purifying our products we find out that the concentration of DNA that we obtained is too low (about 5ng/ul) maybe for an experimental error in setting the PCR program. For this reason, our team is going to redo the PCR reaction again.",
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"tags" : "j23100_YF1_FixJ_FixK2_CI_Plambda_amilCP"
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"tags" : "SAMsynthetase"
}
}

Revision as of 12:27, 19 August 2013

{ "date" : "2013-06-07", "author" : "thomas-emil" "title" : "Second SAMsynthase extraction attempt - TOO SAD", "content" : "Since we didn't obtained our product using the Phusion PCR, we tried again to amplify the SAMsynthase gene from an extract of E.coli genomic DNA (strain MG1655). In order to do that we exploited the same primers used in our previous attempt (see 06-06-2013). For the protocol used see the RBC Taq PCR protocol.. When the reaction finished, we tested the presence of the aplificate product througt an electrophoresis analisys (adding 2 µl of LD for 10 µl of DNA). Results:

Gel Image
As you can see from our gel image, our product is present in both reactions (TAQ and TAQ+Phusion). After purifying our products we find out that the concentration of DNA that we obtained is too low (about 5ng/ul) maybe for an experimental error in setting the PCR program. For this reason, our team is going to redo the PCR reaction again.", "tags" : "SAMsynthetase" }

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