From 2013.igem.org
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| | { | | { |
| - | "date" : "2013-07-31", | + | "date" : "2013-07-17", |
| - | "author" : "thomas", | + | "author" : "fabio-bruno", |
| - | "title" : "pSpac + GFP cloning!!!", | + | "title" : " blue and red light sensors!!", |
| - | "content" : "<html>Today I started a new cloning in order to obtain GFP in the vector pSBBs0K-Pspac (<a href=\"http://parts.igem.org/Part:BBa_K823026\">BBa_K823026</a>). This construct will be useful to Emil in order to characterize Pspac at difference concentration of inducer (IPTG). I started performing a PCR on the GFP following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Phusion-PCR\">Phusion PCR protocol</a>. I used BB_fwd and BB_rev primers to obtain an amplification of the insert. This because the GFP backbone (<a href=\"http://parts.igem.org/partsdb/get_part.cgi?part=BBa_E0840\">BBa_E0840</a>) had the same antibiotic resisistance of our destination vector BBa_K823026 (Amp). I obtain a PCR product with a concentration of 146 ng/µl, confirmed by electrophoresis analysis.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel image|<html><center><img src=\"https://static.igem.org/mediawiki/2013/5/5c/Tn-2013_gel_E0840_PCR.jpg\" width=\"450px\" /></center></html>}}<html> I continued then with the restriction digestions following <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">this protocol</a>. I digested BBa_K823026 using SpeI and PstI, and PCR product with XbaI and PstI. <br/>The two digestion products were then purified and quantified: GFP PCR 60 ng/µl, BBa_K823026 19 ng/µl. The last step was then the ligation that was performed following <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Ligation\">this protocol</a>. The ligation products were then plated on CM plates. </html>", | + | "content" : "<html> today we made 2 inocula for each one of the 4 parts we transformed!! We’re ready to see what’s next! </html>", |
| - | "tags" : "pSpac-GFP" | + | "tags" : " YF1_FixJ - FixK2- ho1_PcyA_cph8-OmpR" |
| | } | | } |
Revision as of 08:33, 3 October 2013
{
"date" : "2013-07-17",
"author" : "fabio-bruno",
"title" : " blue and red light sensors!!",
"content" : " today we made 2 inocula for each one of the 4 parts we transformed!! We’re ready to see what’s next! ",
"tags" : " YF1_FixJ - FixK2- ho1_PcyA_cph8-OmpR"
}
"
