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A recent study examining mouse kidneys took a similar long-term culturing tactic using a transgenic line in which the Myh9 (myosin heavy chain 9) gene was disrupted with a GFP cassette, marking several differentiated cell types in nephrons, as well as the interstitium [ 149].
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Because RRN5 gene is an essential gene, a helper plasmid pRRN5 harboring wild type RRN5 gene expression cassette marked with URA3 gene was introduced into SH6471 strain prior to RRN5 gene disruption.
The subsequent recombination event deletes the region of interest but leaves an FRT- attP−flanked RMCE cassette marked by the w + and y + visible markers.
The version of the RFP-GFP cassette marked with hphMX instead of kanMX (in strains JRY9729 and JRY9730) was constructed by transforming strains containing the kanMX-marked RFP-GFP cassette with PCR product amplified from pAG32 (Goldstein and McCusker, 1999) using the P TEF fwd and T TEF rev primers.
This can be performed by including only one needle-biopsy per cassette, or marking multiple individual cores in one cassette for instance by inking.
And it definitely is hidden – only the small logo of a cassette tape marks the spot.
LoxP sequences flanking the neomycin selection cassette are marked by triangles.
Acute myeloid leukemia cells were transduced with a bicistronic lentivector that encodes the human IL-12 cDNA as a fusion, as well as a LNGFR (ΔLNGFR)/mutant thymidylate kinase cassette as a marking and cell-fate control element.
These experiments are facilitated by the tetracycline resistance cassette used to mark the sigS mutant.
The 9X-Myc epitope tag was amplified from a TRP1 marked cassette (Knop et al. 1999) using DDO1462/-1463 and transformed into wild-type and tdna mutants.
The first two series were first released on audio cassette and CD in 1988, marking the tenth anniversary of the first broadcast of the first episode.
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com