Haynes Lab:Notebook/Engineering PC-TFs/2013/10/23: Difference between revisions
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==October 23, 2013== | ==October 23, 2013== | ||
'''Transfection of KAH126''' | '''Transfection of KAH126''' | ||
{| {{table}} cellspacing="7" width=700px | |||
|- | |||
| Plate-Well || Plasmid || DNA || Volume|| dH2O|| Lipo || PLUS reagent || <u>Opti-MEM (total)</u> | |||
|- | |||
| 1-2 || KAH126 || 1.5 μg || 23 μL > 20 μL || 0μL|| 2.5μL|| 7.5 μL || 570 μL | |||
|} | |||
# Prepare DNA-Lipofectamine complexes (600 μl per sample) as follows: | # Prepare DNA-Lipofectamine complexes (600 μl per sample) as follows: | ||
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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. | |||
Latest revision as of 23:30, 26 September 2017
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October 23, 2013Transfection of KAH126
Cell Culture of KAH126 Grow liquid culture of KAH126
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