Team:UNIPV-Pavia/Protocols/PlasmidExtraction

From 2008.igem.org

(Difference between revisions)
(New page: {| style="color:#000000;background-color:#ffffff;" cellpadding="3" cellspacing="1" border="3" bordercolor="#3128" width="80%" align="center" !align="center"|30px [[Team...)
 
(13 intermediate revisions not shown)
Line 14: Line 14:
<br>
<br>
 +
 +
<h1>The protocols we used</h1>
 +
 +
*[[Team:UNIPV-Pavia/Protocols/Lb|LB medium preparation]]
 +
*[[Team:UNIPV-Pavia/Protocols/Resuspension|Plasmid resuspension from IGEM paper spots]]
 +
*[[Team:UNIPV-Pavia/Protocols/Transformation|Transformation]]
 +
*[[Team:UNIPV-Pavia/Protocols/PlasmidExtraction|Plasmid extraction]]
 +
*[[Team:UNIPV-Pavia/Protocols/Digestion|BioBrick digestion with restriction enzymes]]
 +
*[[Team:UNIPV-Pavia/Protocols/GelExtraction|DNA gel extraction]]
 +
*[[Team:UNIPV-Pavia/Protocols/Precipitation|DNA precipitation with sodium acetate]]
 +
*[[Team:UNIPV-Pavia/Protocols/AntarcticPhosphatase|Antarctic Phosphatase]]
 +
*[[Team:UNIPV-Pavia/Protocols/Ligation|Ligation]]
 +
*[[Team:UNIPV-Pavia/Protocols/Pcr|PCR]]
 +
<h1>Plasmid extraction</h1>
<h1>Plasmid extraction</h1>
-
<h2>QIAGEN Plasmid Mini Kit</h2>
+
''(Estimated time: 3 hours if you use QIAGEN Plasmid Mini Kit; 1 hour if you use QIAGEN QIAprep Spin Miniprep Kit)''
-
''(estimated time: 3 hours)''
+
<br>
<br>
<br>
<br>
'''Materials needed:'''
'''Materials needed:'''
-
*'''Overnight mini-culture
+
*'''5 ml or 9 ml (for higher yields) overnight mini-culture
-
*'''QIAGEN Plasmid Mini Kit'''
+
*'''QIAGEN Plasmid Mini Kit or QIAGEN QIAprep Spin Miniprep Kit'''
<br>
<br>
-
*You can find the detailed protocol at: [http://www1.qiagen/HB/QIAGENPlasmidPurification_EN http://www1.qiagen/HB/QIAGENPlasmidPurification_EN]*Incubate on ice for 30-45 min.
+
*Follow QIAGEN standard protocol: [http://www1.qiagen.com/literature/render.aspx?id=369&tp=4 QIAGEN Plasmid Mini Kit protocol] or [http://www1.qiagen.com/literature/render.aspx?id=370&tp=4 QIAGEN QIAprep Spin Miniprep protocol]
-
*Heat shock: 42°C for 1 min.
+
*We used QIAGEN Plasmid Mini Kit in the first 2 weeks of our activity to perform miniprep, when we didn't have QIAGEN QIAprep Spin Miniprep Kit at our disposal. Then, we always used QIAGEN QIAprep Spin Miniprep Kit for its higher yield.
-
*Put transformed TOP10 tube on ice and then add 250 µl SOC medium.
+
-
*Incubate 2 hours at 37°C, 220 rpm.
+
-
*Centrifuge 10 min, 1200 rpm.
+
-
*Remove 150 ul of bacteria-free supernatant.
+
-
*Plate the remaining part of solution (resuspending the bacteria) on a proper agar plate.
+
-
*Incubate overnight at 37°C.
+
<br>
<br>

Latest revision as of 12:52, 1 July 2008

Home.jpg Home Unipv logo.jpg The Team And.jpg The Project Safety.jpg Biological Safety Dna.png Parts Submitted to the Registry
Laptop.jpg Dry Lab Pipette.jpg Wet Lab Math.gif Modeling Note.jpg Protocols Notebook.gif Activity Notebook


The protocols we used


Plasmid extraction

(Estimated time: 3 hours if you use QIAGEN Plasmid Mini Kit; 1 hour if you use QIAGEN QIAprep Spin Miniprep Kit)

Materials needed:

  • 5 ml or 9 ml (for higher yields) overnight mini-culture
  • QIAGEN Plasmid Mini Kit or QIAGEN QIAprep Spin Miniprep Kit


  • Follow QIAGEN standard protocol: [http://www1.qiagen.com/literature/render.aspx?id=369&tp=4 QIAGEN Plasmid Mini Kit protocol] or [http://www1.qiagen.com/literature/render.aspx?id=370&tp=4 QIAGEN QIAprep Spin Miniprep protocol]
  • We used QIAGEN Plasmid Mini Kit in the first 2 weeks of our activity to perform miniprep, when we didn't have QIAGEN QIAprep Spin Miniprep Kit at our disposal. Then, we always used QIAGEN QIAprep Spin Miniprep Kit for its higher yield.


Retrieved from "http://2008.igem.org/Team:UNIPV-Pavia/Protocols/PlasmidExtraction"