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The difference vector pool will guide the mutation search in the next generation once only.
bIncriminated vector pool.
These data suggest that the size and complexity of a vector pool can be increased only at the expense of losing more slow growing mutants.
In this example, transmission by Cx. vomerifer occurred < 24 h after exposure of the trap, and the vector pool had a titer of 5.8 log10 PFU/pool.
Three batches of schizonts were transfected with the same vector pool and each injected intravenously into a different Balb/c mouse.
In each of two replicate experiments, the same 22 barcodes from the vector pool were robustly detected and yielded nearly identical growth curves.
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This is carried out using vector pools expressing ~10,000 unique gRNA sequences, cloned using the synthesised oligonucleotide libraries of up to 200 bp [ 25].
Once the vector-specific integration rates and fitness measurements are available for the entire vector resource, it will be possible to create more bespoke vector pools that are better matched for integration rates and fitness outcomes.
The use of vector pools to phenotype mixtures of mutants is particularly suited to P. berghei, where a library of cloned mutants would be of limited value, because the absence of an in vitro culture system for blood stages means that each cloned mutant has to be generated and propagated in mice.
To reduce nonspecific binding to Escherichia coli (XL1-Blue MRF') (Stratagene) and phage vector, diluted pool was preadsorbed three times on nonrecombinant phage plaques.
7 ml (pools of 10) or 3.5 ml (pools of 20) of each vector were pooled (70 ml) for ultracentrifugation in two 35 ml-tubes, the rest of the supernatant was used to prepare individual vectors.
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