Haynes Lab:Notebook/Engineering PC-TFs/2013/10/23: Difference between revisions
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| Plate-Well || Plasmid || DNA || Volume|| dH2O|| Lipo || PLUS reagent || <u>Opti-MEM (total)</u> | | Plate-Well || Plasmid || DNA || Volume|| dH2O|| Lipo || PLUS reagent || <u>Opti-MEM (total)</u> | ||
|- | |- | ||
| 1- | | 1-2 || KAH126 || 1.5 μg || 23 μL > 20 μL || 0μL|| 2.5μL|| 7.5 μL || 570 μL | ||
|} | |} | ||
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# Incubate cells at 37°C in a CO<sub>2</sub> incubator | # Incubate cells at 37°C in a CO<sub>2</sub> incubator | ||
# (Optional) To reduce toxicity, after 5-6 hours wash cells once with warm 1xPBS to remove excess DNA-Lipo complexes. Add fresh antibiotic-free growth medium. | # (Optional) To reduce toxicity, after 5-6 hours wash cells once with warm 1xPBS to remove excess DNA-Lipo complexes. Add fresh antibiotic-free growth medium. | ||
'''Cell Culture of KAH126''' | |||
''Grow liquid culture of KAH126'' | |||
*Label 15 ml sterile culture tubes. Fill each tube with 2 ml LB growth medium with Amp. | |||
*Use sterile pipette tip to touch bacterial streak and put the tip into LB medium. | |||
*Grow the cultures for 10-12 hrs. | |||
Revision as of 09:35, 25 October 2013
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October 23, 2013Transfection of KAH126
Cell Culture of KAH126 Grow liquid culture of KAH126
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