as templates, the structures of either SiCYS-N or SiCYS-C showed similarity as those using Orzyacystatin I as templates (data not shown).
Each 25 μl PCR reaction mixture contained 1 μl of each primer (final concentration 800 nM), 2 μl of 12.5 mM dNTPs, 3 μl of 25 mM MgCl2, 0.125 μl of AmpliTaq Gold (5 U. μl-1), 2.5 μl of 10× SYBR Green buffer (SYBR Green Core Reagents kit, Applied Biosystems), 10.375 μl H2O and 5 μl of template containing either 10 ng of cDNA or diluted plasmid DNA containing the relevant cloned PCR product.
If the 3' end of either template is not within the region of similar sequence, then no 3' end will hybridize to the other template and synthesis does not lead to recombination.
O6meG in the template may direct incorporation of either C or T. Thus, TDG may remove T and give the BER system a second chance to incorporate C opposite of the miscoding O6meG-residue.
The bi-species control experiments show that very short regions of sequence identity are sufficient to mediate PCR recombination, the result of either template exchange by Taq polymerase [44] or premature extension termination within the repeat and subsequent illegitimate priming by incompletely extended products [41].
In addition we find that its accurate replication is facilitated by template switching, as loss of either RAD5 or MMS2 leads to a significant decrease in accurate replication.
Total RNA was reverse transcribed into first- and second-strand cDNA, after which first-strand cRNA was constructed using the second-strand cDNA as a template, in the presence of either Cy3-CTP or Cy5-CTP.
We could form any type of vessels, filters, templates, and others made of either plastics or gel through light-induced polymerization by controlling the light pattern generated by DMD (Fig. 2b-d).
The template DNA was 5.0 μL of either a boiled cell suspension or 50 ng genomic DNA.
