User:Karmella Haynes/Notebook/PcTF Genomics/2015/10/20: Difference between revisions
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* | * Luc/CFP swap - Colony growth | ||
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''' | '''Luc/CFP swap - Colony growth'''<br> | ||
* 10/19/15 - Ben Nyer - Gal4EED/luc HEK293 cells were co-transfected with (1) HPK-CFP gene flanked with homology arms that match luciferase site [https://benchling.com/s/IxMj4FhL/edit DBN007_pSB1A3] and (2) Cas9/gRNA34 plasmid | |||
* Today: plate cells at low concentration to try to get CFP-positive colonies | |||
Images of transfected cells: | |||
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[[Image:someimage.jpg|400px]] | |||
Dilution procedure: | |||
# Wash with PBS and trypsinize in 0.5 mL trypsin medium | |||
# Resuspend by adding 4.5 mL DMEM complete (+0.5 μg/mL puro) ≈ 200,000 cells/mL (assuming 1 million cells in 1 well, 6-well dish...there are probably fewer) | |||
# Add 0.5 mL resuspended cells to 10 mL medium in 10 cm plate ≈ 10,000 cells/mL | |||
# Add 50 μL resuspended cells to 10 mL medium in 10 cm plate ≈ 1,000 cells/mL | |||
Plates: | |||
# Well 2 | |||
Revision as of 19:50, 20 October 2015
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10/20/15
Luc/CFP swap - Colony growth
Images of transfected cells: Dilution procedure:
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