Team:UNITN-Trento/Notebook/Labposts/06/26

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(Created page with "{ "date" : "2013-06-13", "author" : "caterina-michele", "title" : " A Long Day", "content" : " <html>We took 4 inocula of <b>pSB1C3 </b> and we performed the purification (Wi...")
 
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{
{
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"date" : "2013-06-13",
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"date" : "2013-06-14",
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"author" : "caterina-michele",
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"author" : "thomas",
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"title" : " A Long Day",
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"title" : "EFE extraction and transformation Test!",
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"content" : " <html>We took 4 inocula of <b>pSB1C3 </b> and we performed the purification (Wizard Plus SV Miniprep DNA purification system). Then we performed a digestion with EcoRI and PstI of this 4 minipreps, SAM (another time because we were not satisfied of the previous results!) and two pSB1C3. After the digestion we did a gel to understand if it was our lucky day. As you can see from the picture, evidently not! </html> {{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel Image|<html><img id=\"post_img\" src=\"https://static.igem.org/mediawiki/2013/0/0a/Tn-20130613-Super_Gel_screening_8_cose.jpg\" width=\"500px\"/> </html>}}<html>In the afternoon we prepared two PCR reactions: one for J45700 (from the miniprep = it’s the complete device!) and one to linearize and to amplify the pSB1C3 vector. Both the reactions failed and we were too disappointed to take a picture of the gel.Finally we made another digestion for J45319, J45119 and Cate’s pSB1C3  with EcoRI and PstI. </html>",
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"content" : "Since we received the Ethylen Forming Enzyme gene from Genescript company, we proceeded with the extraction and the transformation test.We resuspended the 4 ug of DNA in 40 ul of sterilized water (obtaining a 100 ng/ul stock solution). After that, we transformed 200 ul of NEB10 competent cells with 1ul of EFE DNA.Finally we plated them in two 50 ug/ml Amp LB-Agar Petri dishes.'''Results:'''{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Plate Image|<html><img id=\"post_img\" src=\"https://static.igem.org/mediawiki/2013/4/42/Tn-20130614-plate_TP.JPG\" width=\"500px\"/></html>}}As you can see from the image, we obtained many colonies. We then inculated x5 colonies in 5ml LB with Ampicillin.",
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"tags" : "PchBA-PchA-PchB"
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"tags" : "EFE"
}
}

Latest revision as of 07:47, 3 October 2013

{ "date" : "2013-06-14", "author" : "thomas", "title" : "EFE extraction and transformation Test!", "content" : "Since we received the Ethylen Forming Enzyme gene from Genescript company, we proceeded with the extraction and the transformation test.We resuspended the 4 ug of DNA in 40 ul of sterilized water (obtaining a 100 ng/ul stock solution). After that, we transformed 200 ul of NEB10 competent cells with 1ul of EFE DNA.Finally we plated them in two 50 ug/ml Amp LB-Agar Petri dishes.Results:

Plate Image
As you can see from the image, we obtained many colonies. We then inculated x5 colonies in 5ml LB with Ampicillin.", "tags" : "EFE" }

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