Although compartmentalized self-replication (CSR) and compartmentalized partnered replication (CPR) are powerful tools for directed evolution of proteins and gene circuits, limitations remain in the emulsion PCR process with the wild-type Taq DNA polymerase used so far, including long run times, low amounts of product, and false negative results due to inhibitors.
This may be due to optimal levels of expression already achieved with the native gene and limitations of the chloroplast protein synthetic machinery other than codon usage.
When compared with microarray technology that requires previous knowledge of genes, the limitations to detecting unknown genes are not encountered in MPSS [ 48, 49].
When compare with cDNA microarray technology that requires previous knowledge of genes, the limitations to detect unknown genes was not encountered in MPSS [ 32, 39].
More and more evidences have shown that conventional gene-level analysis methods seeking biomarkers or differential genes encountered limitations and difficulties from both statistical and biological sides [46] [47].
As Xist is not a protein-coding gene, this limitation is removed.
Mapping SNPs, identification of the smallest P value of each gene, and limitation of this approach was described in our previous study [11].
However, we expected that favored alleles would generally be located within extended and shared haplotypes [13], [27] that encompass larger regions, partially compensating for the gene-centric limitations of the dataset.
When MG is applied to very short sequences containing one or two partial genes, these limitations are not significant.
This is probably a consequence of inadequate pollinator visitation to small populations, resulting in strong gene flow limitation [ 2, 54].
