其中最常见的计算方法是使用“Melting Temperature(Tm) Calculator”软件,该软件可以根据输入的DNA序列信息和实验条件进行DNA解链温度的计算。 在实验过程中,通常会先将DNA样品加热至高温,使DNA解链,然后逐渐降温,记录DNA的回复过程。通过分析回复过程中的吸光度变化,可以确定DNA解链温度。 总之,DNA解链温度的计算具有...
The ions were selected for CID analysis and the elucidation of the CID fragmentations of the candidate oligodeoxynucleotide sequence was done with the aid of the Mongo Oligo Mass Calculator (v. 2.06) from The RNA Institute, State University of New York at Albany (http://mods.rna.albany.edu/...
The Gene Designer can also be used to design oligonucleotides. To assist with this, a real-time Tm calculator can be positioned in the Sequence View and dragged until a preferred location, length and melting temperature is found. The DNA melting temperature calculation is performed using the near...
3b). This was in agreement with a thermodynamic analysis using the Oligonucleotide Properties Calculator (Supplementary Fig. 3c, Methods, Eqs. (3)–(5))28,29,30. DORIS increases density and capacity limits One potential advantage of room temperature separations of files is that the double-...
Simplified workflow—Buffer formulated for primer annealing at 60°C (no need for a Tm calculator) Increased PCR success—Robust amplification of GC-rich targets, DNA of suboptimal purity, and long sequences Enabled automation—High specificity and benchtop stability for 24 hours af...
Data from the replicate standard curves were analysed in RStudio (v1.2.5042) running R (v4.0.0) using the Generic qPCR Limit of Detection (LOD) / Limit of Quantification (LOQ) calculator R Script (Klymus et al., 2020, Merkes et al., 2019). A three-parameter logistic function with ...
The genome of the obligate biotrophic phytopathogenic barley powdery mildew fungus Blumeria hordei is inflated due to highly abundant and possibly active transposable elements (TEs). In the absence of the otherwise common repeat-induced point mutation tr
(Online Resource TableS2). This pair also had a melting temperature difference < 5 °C, which is within the recommended limit (Ye et al.2012; see tmcalculator.neb.com) and allows for efficient annealing during a PCR. This difference was better than either of the other universal primer...
but often between about 0.5 nM and 5 nM. Betaines and lower tetraalkyl ammonium salts are examples of isostabilizing agents. Denaturing agents are compositions that lower the melting temperature of double stranded nucleic acid molecules by interfering with hydrogen bonding between bases in a double-...
15 to 20 nucleotides long, on both ends so that the melting temperature of the homologous sequences will not be higher than 65° C. to allow complete denaturation at 75° C. The reagents are combined on ice in