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PCR analysis showed that 3 clones were free of vectors, but clone NR2-d20 still retained residual amount of vectors (Fig. 2G).
The recently developed iPS cells free of vectors and transgene sequences may provide a solution for this problem [ 130- 134].
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Yu, J. et al. Human induced pluripotent stem cells free of vector and transgene sequences.
Moreover, our oriP-EBNA1-based CRISPR/Cas9 system was free of vector in both genetically modified miPSCs and human cells.
For microinjection the transgene was isolated free of vector sequences by double digestion with Sall and Xbal, and the transgene was purified by agarose gel electrophoresis.
Sequence trimming was conducted using LUCY – a program developed at TIGR [ 33] with a trimming standard of an overall base call error rate of <1%, free of vector- and E. coli sequences, and a trimmed sequence read length of > 100 bp.
By the end of the epidemic in January 2002, a broad hospitalization policy was established in areas free of vector, Aedes aegpyti mosquitoes, (all febrile and dengue suspected case-patients were hospitalized or treated at home with daily visits by the family doctor) (13 ).
The insert DNA segment free of contaminating vector sequences can be used as a hybridization probe and for insertion into a new vector since suitable cohesive termini are generated from the retained poly dA) and poly dT) tails by an appropriate exonuclease.
Consisting of only one active transcription unit and the S/MAR, it resists silencing as it is free of prokaryotic vector parts and drug selection markers.
Finally, we examined whether these four successfully engineered miPSCs (NR1, NR2, NR3, NR5) were free of targeting vector.
Subsequently, the cassette downstream of the promoter can be replaced by another coding unit based on the same set of flanking heterospecific FRT sites and free of nonessential vector components ("recombinase-mediated cassette exchange" (RMCE)) exploiting the potential of yeast FLP recombinase (Qiao et al. 2009).
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