Team:EPF-Lausanne/Protocol/GelExtraction

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in length between the wanted fragment and the closest-sized fragment should
in length between the wanted fragment and the closest-sized fragment should
be more than 200bp). These fragments are often obtained after a
be more than 200bp). These fragments are often obtained after a
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[[Team:EPF-Lausanne/Protocol/Digestion|digestion]] .
+
[[Team:EPF-Lausanne/Protocol/RestrictionSiteDigestion|digestion]].
Start with a lot of DNA (at least 2 ug of DNA to get reasonable yields), and
Start with a lot of DNA (at least 2 ug of DNA to get reasonable yields), and
-
load a [[Team:EPF-Lausanne/Protocol/Gel|gel]]  
+
load a [[Team:EPF-Lausanne/Protocol/Gel|gel]] with it (if possible, leave an
 +
empty lane on both sides to avoid contamination, these bands get quite big).
 +
When the gel has finished running, put it on a UV light, and locate the correct
 +
band. Cut it out (with a razorblade, try cutting away as much agar as possible
 +
while retaining the DNA) and put it in an Eppendorf (consider using a 2ml one).
 +
Then apply the instructions in your kit.
We used Macherey-Nagel's "Nucleospin® Gel and PCR clean-up" kit.
We used Macherey-Nagel's "Nucleospin® Gel and PCR clean-up" kit.

Revision as of 22:05, 24 September 2012

Protocol: Gel Extraction


A gel extraction is used to select a fragment of DNA of a specific length out of a solution composed of different fragments (ideally the difference in length between the wanted fragment and the closest-sized fragment should be more than 200bp). These fragments are often obtained after a digestion.

Start with a lot of DNA (at least 2 ug of DNA to get reasonable yields), and load a gel with it (if possible, leave an empty lane on both sides to avoid contamination, these bands get quite big). When the gel has finished running, put it on a UV light, and locate the correct band. Cut it out (with a razorblade, try cutting away as much agar as possible while retaining the DNA) and put it in an Eppendorf (consider using a 2ml one). Then apply the instructions in your kit.

We used Macherey-Nagel's "Nucleospin® Gel and PCR clean-up" kit. Its manual can be found here: [http://www.mn-net.com/Portals/8/attachments/Redakteure_Bio/Protocols/DNA%20clean-up/UM_PCRcleanup_Gelex_NSGelPCR.pdf Gel and PCR clean-up Manual]