Genomic Deletions in Mammalian Cell Lines
Type
Collection
...Ensure there are 2 x 10 6 cells per CRISPR pair. Resuspend 2 x 10 6 cells in 100 μl of electroporation solution...Dilute oligos 1:10 in ddH 2 O ( e.g., 1.0 μl annealed oligos + 9.0 μl ddH 2 O to yield a concentration...), 50-100 ng gDNA, and H 2 O up to 20 μl. Use the primers designed in step 2 above. Conduct PCR for “non-deletion...supplemented with 2% penicillin/
streptomycin and 1% L-glutamine, which is used for MEL cells. However, transient...expression construct. Electroporate cells with 250 volts for 5 msec in a 2 mm cuvette using an electroporation...designed based on testing the bulk sorted cells. Run samples on 2% agarose gel at 10 V/cm using 1x Tris-acetate-EDTA...frequency of indels formed on non-deleted alleles 2 , which are likely to cause frameshift mutations leading...