From 2013.igem.org
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| | { | | { |
| - | "date" : "2013-08-07", | + | "date" : "2013-08-06", |
| - | "author" : "thomas", | + | "author" : "bruno-viola-caterina-gabriele-thomas", |
| - | "title" : "pXil + GFP transformation in B. subtilis", | + | "title" : "Surrender now or prepare to fight, Meowth! That's right.", |
| - | "content" : "<html> In order to transform <i>Bacillus</i> with pXyl (BBa_K823024) + GFP (BBa_E0840), reamplified and screened <a href=\"https://2013.igem.org/ wiki/index.php?title=Team:UNITN-Trento/ Notebook# tn- post- 2013-08-02-thomas\"> the 2nd of August</a> , I prepared an inoculum in 2 ml of MN completed medium (<a href=\"https: //2013.igem.org/Team:UNITN-Trento/Protocols#subtilis-transformation\">see protocol</ a> ). While I was waiting that O.D.600 reached 0.5, I proceeded digesting 2,5& micro; g of sample 2 and sample 3 of the construct pXyl + GFP using ScaI as restriction enzyme. I did this in order to linearize the plasmid and allow his integration in < i> B. subtilis</ i> genome. 0.5µg of each sample were then run on an agarose gel for screening.</ html> {{:Team:UNITN- Trento/ Templates/Styles/Spoiler|Gel image|<html>< center>< img src=\"https:/ /static. igem.org/ mediawiki/ 2013/9/90/Tn-2013_gel_pXyl%2BGFP_ScaI_digested.JPG\" width=\"450px\" /></ center></ html> }}< html> As you can see from the picture, there are two not expected bands: one at height of 4500 bp and one at height of about 1800 bp. I can't explain this two bands since ScaI should digest only at only one sites on the construct.. However there's also a band at the height between 5000 and 6000 bp as expected (pXyl+GFP = 5782 bp) so I transformed it into < i> bacillus</ i> anyway.</html>", | + | "content" : "<html> Today we made 5 different competent cells following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/ Protocols# Competent- cells- transformation\"> transformation protocol</a>:< br/> - NEB10& beta;< br/> - TB1< br/> - TOP10< br/>- JM109<br/> - BL21< br/>< br/ >Then we trasformed in these cells R0010-K592009 in psB1C3 and K1065300. <br/ ><br/ ><br/>< br/>< br/>< br/>< br/>< br/></html>", |
| - | "tags" : "pXyl-GFP-B. subtilis" | + | "tags" : "cells competent" |
| | } | | } |
Latest revision as of 09:05, 3 October 2013
{
"date" : "2013-08-06",
"author" : "bruno-viola-caterina-gabriele-thomas",
"title" : "Surrender now or prepare to fight, Meowth! That's right.",
"content" : "Today we made 5 different competent cells following the transformation protocol:
- NEB10β
- TB1
- TOP10
- JM109
- BL21
Then we trasformed in these cells R0010-K592009 in psB1C3 and K1065300.
",
"tags" : "cells competent"
}
"
