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Ethanol precipitation of small DNA fragments: Difference between revisions
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Ethanol precipitation of small DNA fragments (view source)
Revision as of 19:46, 30 May 2005
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This protocol is for a simple ethanol precipitation of small fragments. This protocol was used to (partially) purify a DNA fragment containing a ribosome binding site (~40 bp) during 3A [http://rosalind.csail.mit.edu/r/parts/htdocs/Assembly/index.cgi assembly]. The fragment was generated via restriction digest and it was used in a ligation reaction. Note that this protocol simply concentrates your sample and removes enough salts/enzymes for ligation to be successful. All DNA fragments from your digest will still be present in your pellet. These residual DNA fragments do not matter for 3A assembly which selects against incorrect ligation products. | This protocol is for a simple ethanol precipitation of small fragments. This protocol was used to (partially) purify a DNA fragment containing a ribosome binding site (~40 bp) during 3A [http://rosalind.csail.mit.edu/r/parts/htdocs/Assembly/index.cgi assembly]. The fragment was generated via restriction digest and it was used in a ligation reaction. Note that this protocol simply concentrates your sample and removes enough salts/enzymes for ligation to be successful. All DNA fragments from your digest will still be present in your pellet. These residual DNA fragments do not matter for 3A assembly which selects against incorrect ligation products. | ||
=Materials= | ==Materials== | ||
*Absolute Ethanol (200 proof) | *Absolute Ethanol (200 proof) | ||
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*-80°C freezer | *-80°C freezer | ||
=Procedure= | ==Procedure== | ||
#Add 2 volumes ice cold absolute ethanol to sample. <br> Generally the sample is in a 1.5 mL eppendorf tube. I recommend storing the absolute ethanol at -20°C. | #Add 2 volumes ice cold absolute ethanol to sample. <br> Generally the sample is in a 1.5 mL eppendorf tube. I recommend storing the absolute ethanol at -20°C. | ||
