From 2013.igem.org
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| | { | | { |
| - | "date" : "2013-06-12", | + | "date" : "2013-06-11", |
| - | "author" : "Gabriele", | + | "author" : "emil-fabio", |
| - | "title" : "Ligation and transformation", | + | "title" : "", |
| - | "content" : "<html>Ligation of SAMsynthetase and pSB1C3 with and without RFP, so I prepared four samples:<br/>1) pSB1C3 with RFP (Ctrl_1)<br/>2) pSB1C3 with RFP + SAMsynthetase (1:2)<br/>3) pSB1C3 without RFP (Ctrl_2)<br/>4) pSB1C3 without RFP (1:2)<br/>The samples were prepared and ligation was performed following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Ligation\">ligation protocol</a>.<br/><br/></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Ligation parameters|<html><ul><li>Plasmid concentration: 27.8ng/µl</li><li>Plasmid length: 2070bp</li><li>Insert concentration: 13.2ng/µl</li><li>Insert length: 1155bp</li><li>Used plasmid: 250µl</li><li>Volume of reaction: 35µl</li><li>Buffer concentration: 10X</li></ul></html>}}<html><br/>Transformation in NEB10β cells was performed following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Competent-cells-transformation\">transformation protocol</a>.Plates in incubator ON at 37°C static, more than 16 hours.</html>", | + | "content" : "<html><h5>Inoculum of three B.subtilis backbone</h5>We have done the inoculum of three Bacillus-specific backbone (previously transformed in NEB and plated on Ampicillin LB Agar) in 19 50ml Falcon- 7 with the part BBa_K823023- 6 with the part BBa_K823022- 6 with the part BBa_K823027The Falcons were put in the incubator at 37 °C o/n.<h5>Digestion of SAMsynthetase and psB1C3</h5>We have digested the SAMsynthetase gene (previously amplificated and purificated)and the linerarized vector psB1C3 with the enzyme Xba1 and Pst1 exploiting the same protocol. Then we have incubated the mix at 37C o/n.</html>", |
| - | "tags" : "SAMsynthetase" | + | "tags" : "SAMsynthetase-pSBBs1C-pSBBs4S-pSBBs2E" |
| | } | | } |
Latest revision as of 07:41, 3 October 2013
{
"date" : "2013-06-11",
"author" : "emil-fabio",
"title" : "",
"content" : "
Inoculum of three B.subtilis backbone
We have done the inoculum of three Bacillus-specific backbone (previously transformed in NEB and plated on Ampicillin LB Agar) in 19 50ml Falcon- 7 with the part BBa_K823023- 6 with the part BBa_K823022- 6 with the part BBa_K823027The Falcons were put in the incubator at 37 °C o/n.
Digestion of SAMsynthetase and psB1C3
We have digested the SAMsynthetase gene (previously amplificated and purificated)and the linerarized vector psB1C3 with the enzyme Xba1 and Pst1 exploiting the same protocol. Then we have incubated the mix at 37C o/n.",
"tags" : "SAMsynthetase-pSBBs1C-pSBBs4S-pSBBs2E"
}
"
