However, the final targeted allele should be free of selection cassettes and screening for correct removal of selection cassettes can be a laborious task.
After PCR screening for correct homologous recombination of the 5′ (Hotstar Taq: Qiagen, West Sussex, UK) and 3′ flank (Long Range Taq: Fermentas, York, UK) only one clone was found to be positive.
The high efficiency of our approach allowed us to screen for correct modification of the loci by simple genomic polymerase chain reaction (PCR) of a limited number of individual fly lines.
Subcloning efficiency DH5alpha bacteria (Life Technologies, Grand Island, NY, USA) were transformed with the ligated vector and colonies screened for correct insertion of hCXCR7 using differential enzyme digestions, followed by verification using sequencing analysis.
This screens for correct recombination events of the PCR amplicon with the target plasmid.
The landing site is designed such that correct transgene integration results in a specific change of fluorescence, allowing to screen for correct genomic integration and therefore to preselect for transgenic founder fish.
The resulting clones were screened for correct orientation and for the regeneration of the SalI site.
PCR screens of the correct size were treated with exonuclease I and shrimp alkaline phosphatase [ 34] and sequenced at the University of Hawaii's Advanced Studies in Genomics, Proteomics, and Bioinformatics Facility.
This was subsequently screened for correct orientation.
Men were tested for syphilis if they sought medical care for symptoms or accepted screening, with a likelihood of correct diagnosis dependent on test sensitivity [ 20, 44].
Since the mid-1990s, maternal serum testing and, later, measurement of fetal nuchal translucency, were successful screening tests, and antenatal screening has achieved higher rates of correct predictions and higher coverage year on year.
