Lactococcus transformation: Difference between revisions

Lactococcus transformation (view source)

12 bytes removed , 5 April 2011

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 ===Electro-Transformation===
 .  Add 5µl of DNA and electroporate at 1200 volts (1mm Cuvettes). <br>
 .  Add 900µl ice cold M17+ and keep on ice for 10 mins. <br>
-::*(0.5M(.17g)Sucrose + 0.5%(15µl)Glucose+20mM(10µl)MgCl+0.2mM(10µl)CaCl). <br>
+::*To eah ml of M17 add the following: 0.5M(.17g)Sucrose + 0.5%(15µl)Glucose + 20mM(10µl)MgCl<sub>2</sub> + 0.2mM(10µl)CaCl<sub>2</sub>. <br>
 .  Subculture 100µl into 900ul prewarmed M17+ and incubate for 2 hours. <br>
-.  Add entire subculture to GM17 with antibiotic. <br>
+.  Plate with appropriate antibiotic. <br>
-::*1ug/ml Erm for plates. <br>
-::*5ug/ml Erm for culture.
 [[Category:Protocol]]