Because this method will track the target according to the rate of increasing gradient; therefore in comparison to the template matching method, this method is much faster and requires less computational amount, because ignores blindly searching [36].
Moreover, due to high quality normalization, the template also retains high contrast and sharp features in comparison to standard templates.
Thereafter assays results were read at 25°C using an endpoint genotyping programme in a Techne Quantica qPCR thermal cycler (Bibby Scientific Ltd, Stone, UK) in which unknown genotypes were assigned based on fluorescent output in comparison to non-template control wells containing DNA/RNA free H2O.
Figure 1 displays templates for the different modalities in comparison to the MPRAGE template.
Furthermore, addition of Phen-DC3 did not affect significantly the RNA synthesis on MutG4 in comparison to the WT template.
The nhr-25 :: 3xFLAG oligo appeared to be a poor repair template, as it yielded lower knock-in efficiency in comparison to the nhr-25 :: 2×FLAG template, and complex insertions were observed in two nhr-25 :: 3xFLAG candidate knock-in lines.
Under the chosen conditions, the extension for the A-primer was 56%% when paired with the 5mC template in comparison to 18%% when paired with the C template.
This suggests that the metal ions act as bond bridges between functional molecules and silanol groups so that the polymers exhibit very well washing-resistant ability, in comparison to the compact aggregated polymer templates sensitive to washing process [14].
This is evident in a number of studies where reduced digestion with restriction enzymes was observed in nucleosomal templates in comparison to non-nucleosomal ("free") DNA [24], [25], [26], [27].
As demonstrated in Fig. 5, no further repair is seen of the SSB resulting from AP site cleavage within nucleosomal templates, in comparison to efficient repair of the resulting SSB within free DNA.
Interestingly, KlenTaq wild-type exhibits a more sterically crowded environment close to the relevant template site in comparison to KOD exo-.
