( ty PCR applications of 2025 l. ...)TripleMaster PCR System

ty PCR applications of 2025 l.

11.2 Use of PCR additives and co-solvents with TripleMaster PCR System

In most cases, the TripleMaster PCR System produces good results without any additive, with either the Tuning or HighFidelity Buffer. For very GC-rich (>70%) templates or templates containing complex secondary structure, the addition of iso-stabilizing cosolvents such as betaine (0.53 M final concentration), DMSO (25 % final), glycerol (25%), trimethylamine N-oxide (10100 M) or trimethyleamine hydrochloride (10100 M) may improve yield and specificity of PCR. However, the use of destabilizing co-solvents substantially reduces the PCR fidelity and requires a re-optimization of the annealing temperature. The TaqMaster PCR Enhancer, available separately, can be used to stabilize the enzyme for elevated denaturation temperatures (up to 98C) during amplification of GCrich targets. This approach is not feasible in long range PCR.

11.3 Enzyme concentration

Long range applications: 2 U = 0.4 l Other applications: 0.52.5 U = 0.10.5 l 11.4 Concentrations of dNTPs and Mg²⁺

Both buffers of the TripleMaster PCR System provide a final Mg concentration of 2.5 mM which is ideal for most applications. An addition of 25 mM Magnesium Solution to the PCR reactions (in increments of 0.2 mM) is necessary only if the template concentration exceeds 500 ng per 50 l PCR volume and/or the dNTP concentration is higher than 500 M.


'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
Related biology technology :

1. TripleMaster PCR System
2. The TripleMaster PCR System for three different amplifications
3. TripleMaster PCR System: Direct Comparison with Competitors
4. TripleMaster and Perfect gDNA Blood Mini Kit Team Up to Amplify Long gDNA Templates
5. TripleMaster PCR SystemThe Ideal Choice for Long-Range PCR
6. Using the TripleMaster PCR System for Robust Amplification of GC-Rich DNA Templates
7. New Mammalian Two-Hybrid System Detects Protein-Protein Interactions
8. Assess the In Vivo Activation of Signal Transduction Pathways with PathDetect Reporting Systems
9. Generate Adenovirus Vectors in E. coli by Homologous Recombination with the AdEasy Adenoviral Vector System
10. New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
11. Enhanced PCR Cloning System