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Restricted
From 2008.igem.org
Revision as of 14:54, 16 September 2008 by Riachalder (Talk | contribs)
Restricting Plasmids (Double Restriction) From 2008.igem.org
Jess enjoying pipetting buffer into her restriction mixture.We have conducted restrictions in varying concentrations and total volumes; however they all follow the same basic procedure. Below are two of the different restrictions we carried out.
46μl MillQ H2O
10μl 10 x buffer
40μl plasmid sample
2μl enzyme 1
2μl enzyme 2
Total volume = 100μl
Concentrated
10μl MilliQ H2O 3μl 10 X buffer 10μl plasmid sample 1μl enzyme 1 1μl enzyme 2 Total volume = 30μl
The DNA purification kit we use to purify enzymatic reaction mixtures.Incubate solutions for 90 minutes in a 37°C water bath.
If DNA is to be run on a gel and a band cut out, this can be done without purifying the DNA. For all other downstream applications or for storage, solution must be purified to remove the enzymes. Retrieved from "https://2008.igem.org/Restricting_Plasmids_%28Double_Restriction%29"
