Two years ago, company scientists had generated a clone of an endangered wild Asian ox, called a gaur.
Each of the tested single C6 cells was able to generate a clone and subclones in serum-containing medium, which subsequently gave rise to a xenograft glioma in nude mice.
This second mutation must arise during growth under selection (after day 1) since any singly mutant cell plated would generate a clone of only ∼100 cells by the time full selection was imposed insufficient to allow the second mutation.
To establish the different 293T-REx lines, we first generated a clone harboring an inducible shRNA transgene against SCML2 (" shL2" clone); then, we transfected the different transgenic constructs (N3, N3 SCML2AWT and N3 SCML2AΔRBR) into the shL2 clone and new clones harboring both the inducible shRNA and the inducible SCML2 constructs were selected.
In this approach, a library of bacterial artificial chromosome (BAC) clones is mapped onto chromosomes by the use of molecular markers, the aim being to generate a clone-based physical map with a "minimum tiling path".
It has been hypothesized that chromothripsis is a one-off cellular crisis generating a malignant clone in a very short time [ 9, 44].
AFVY-RFP was amplified from 35S::AFVY-RFP (Scheuring et al., 2011) and introduced into pDONR221 (Invitrogen) generating a pEntry clone.
The final PCR product was gel purified and subsequently cloned into an Entry vector, pDONR221 (Invitrogen), to generate a master clone.
Utilizing a commercially acquired GPS®-linker scanning (LS) mutagenesis system, random insertions of 5 amino acids or truncations were introduced into the coding sequence of the plasmid-borne levQ gene, generating a library of clones of levQ mutants.
The 5' RACE clone with the longest 5' UTR region and the cDNA clone P21 were ligated together through a unique PmlI site at # 722 to generate a cDNA clone (Dazap1-C) with a near full-length insert.
