From 2013.igem.org
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| | { | | { |
| - | "date" : "2013-07-20", | + | "date" : "2013-07-31", |
| - | "author" : "thomas", | + | "author" : "fabio", |
| - | "title" : "Digestion of AraC-pBAD + EFE and Venus PCR product", | + | "title" : " the blue ligation: the never ending story !", |
| - | "content" : "<html>This is the part two of <a href=\"https://2013.igem.org/wiki/index.php?title=Team:UNITN-Trento/Notebook#tn-post-2013-07-18-thomas-michele\">this experiment</a>.Since both Venus (BBa_K537006) and AraC-pBAD + EFE were in pSB1C3 vector, I had to do a PCR on Venus using primers Prefix Fwd and Suffing Rev following <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#pSB1C3-linearization-PCR\">this protocol</a>. The PCR product was then confirmed by an electrophoresis analysis (Venus is 729 bp and KAPA universal Ladder was adopted). <br/></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler| Gel Image|<html><center><img src=\"https://static.igem.org/mediawiki/2013/ f/ fe/Tn- 2013_VenusPCR_gel.jpg\" style =\" width: 450px\" ></center></html>}}<html><br/>The PCR product was then purified and quantified at the NanoDrop.Once I obtained Venus without his backbone, I proceeded digesting all the PRC Venus product with NgoMIV and PstI and 2-3 ug of AraC-pBAD + EFE with AgeI and PstI following <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">the PCR product digestion protocol</a>. The digestion products were finally purified, quantified and stored at -20°C.<br/> </html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Quantification Results|<html><center><table class=\"tn-sp-table\"><tr><td>AraC-pBAD + EFE</td><td>33.5 ng/ul</td></tr><tr><td>Venus PCR product</td><td>22.8 ng/ul</td></tr></ table></center></html>}}<html></html>", | + | "content" : "<html> Today massive miniprepping y’all!! This is a wonderful day because one of my miniprep seems toOk!! Sample number 3 from the 3rd ligation doesn’t show the band that correspond to 016 not ligated to the 006 promoter! Tomorrow I’ll need to verify by onetaq pcr.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler| Immagine gel|<html><center><img src=\"https://static.igem.org/mediawiki/2013/ 3/ 3a/Tn- 2013_Fff.jpg\" width=\"450px\" /></center></html>}}<html></html>", |
| - | "tags" : "EFE-Venus" | + | "tags" : "YF1_FixJ-FixK2" |
| | } | | } |
Revision as of 08:46, 3 October 2013
{
"date" : "2013-07-31",
"author" : "fabio",
"title" : " the blue ligation: the never ending story !",
"content" : "Today massive miniprepping y’all!! This is a wonderful day because one of my miniprep seems toOk!! Sample number 3 from the 3rd ligation doesn’t show the band that correspond to 016 not ligated to the 006 promoter! Tomorrow I’ll need to verify by onetaq pcr.
",
"tags" : "YF1_FixJ-FixK2"
}
"
![](\"https://static.igem.org/mediawiki/2013/3/3a/Tn-2013_Fff.jpg\")
