Cellartis iPSC rCas9 Electroporation and Single-Cell Cloning System User Manual
(030619)
takarabio.com
Takara Bio USA, Inc.
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4.
Resuspend the cells in 500 µl per well of pre-warmed DEF-CS SCC medium. Transfer all of the cell
suspension to a newly coated well in a 48-well plate.
NOTE:
To prevent cell loss,
counting the cells at this stage is not recommended.
5.
Tilt the dish backwards and forwards gently to ensure that the cell suspension is dispersed evenly
over the surface, then place it in an incubator at 37°C ± 1°C, 5% CO
2
, and >90% humidity.
X.
Scaling up from the 48-Well Plate
Once the cells have been passaged into a 48-well plate,
DEF-CS GF-3 is no longer needed in the growth
medium
used when changing the media. Prepare “DEF-CS medium for maintenance” by adding DEF-CS GF-1
(dilute 1:333) and GF-2 (dilute 1:1,000) to Cellartis DEF-CS 500 Basal Medium. When the cells are ready to be
scaled up to a 24-well plate, they can be cultured with the Cellartis DEF-CS 500 Culture System (Cat. No.
Y30010).
XI. References
Jinek, M.
et al.
A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity.
Science.
337,
(2012).
Kim, S., Kim, D., Cho, S. W., Kim, J. & Kim, J.-S. Highly efficient RNA-guided genome editing in human cells
via delivery of purified Cas9 ribonucleoproteins.
Genome Res.
24,
1012–9 (2014).
Sander, J.-D. & Joung, J.-K. CRISPR-Cas9 systems for genomic editing, regulation and targeting.
Nat.
Biotechnol.
32,
347–55 (2014).
For information on using the 4D Nucleofector System to electroporate cells, please visit:
http://www.lonza.com/products-services/bio-research/transfection/genome-editing.aspx
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