UsingTaqDNA polymerase has several limitations. It does not possess proofreading activity (i.e., 3' to 5' exonuclease activity), and therefore has a relatively high rate of base misincorporation (error rate). At the end of a typical 30-cycle PCR, a significant proportion of the products ...
Increasing the amount of DNA polymerase from 1 to S U had a strong effect for ExTaq HS, elevating HA-tolerance four times. We also show that the average Cq values of positive reactions may be used as a measure of inhibition effects, e.g., to determine whether or not a dPCR ...
Herein, we present the development of a Taq DNA polymerase-specific repebody, which is a small-sized protein binder composed of leucine rich repeat (LRR) modules, as a thermolabile inhibitor for a precise and accurate gene amplification by PCR. We selected a repebody that specifically binds to...
Total RNA isolation and complementary DNA synthesis Total RNA was extracted using the Nucleospin® RNA Plant kit (Macherey–Nagel, Düren, Germany), starting from 100 mg of finely grinded leaf samples, according to the manufacturer’s instructions. RNA concentration was estimated by reading spectrop...
was ple was heated at picked from agar plates and re-suspended in 30 μ l 96 °C for 5–10 minutes to lyse the cells, and 1 μ l of otfhidsHs2uOspienns0i.o2n mwl atus buesse.dTahse sam- DNA template in 30 μ l PCR reactions using Taq DNA polymerase. ...
FastTaq Premix(with dye) 10 *1 mL 50 mL 1,000 mL FastPfu DNA polymerase 100 U 500 U 1,000 U FastPfu Premix 1 mL 10*1 mL 50 mL 1,000 mL FastPfu Premix (with dye) 1 mL 10*1 mL 50 mL 1,000 mL P6 High-Fidelity polymerase 100 U 500 U 1,000 U P6 High-Fidelity Premix 1...
For each sample, two duplex PCRs were carried out using 5 ng genomic DNA and 0.5 U Taq DNA polymerase (NEB) in a buffer with final concentrations of 50 mM Tris–HCl pH8.8, 12.5 mM ammonium sulphate, 1.4 mM magnesium chloride, 7.5 mM 2-mercaptoethanol, 125 μg/ml BSA and 200 μM ...
minimizing risk of misincorporation. We engineered the active site of 9°N DNA polymerase to improve the efficiency of incorporation of these unnatural nucleotides9. After each cycle of incorporation, we determined the identity of the inserted base by laser-induced excitation of the fluorophores and...
DNA analysis for deletional α-thalassaemia The five common deletions causing α-thalassaemia in Southeast Asia: -α3.7, -α4.2, –SEA, –FIL, –THAIwere characterised using an adapted multiplex-PCR21(Supplementary Table 1). Hot StartTaqDNA polymerase (Qiagen HotStarTaqPlusDNA Polymerase, Hilden,...
aBased on the instruction of manufacturer, the PCR reaction (25 μL) used 0.125 μL of Taq polymerase (1.25 U), 0.5 μL of primers (5 pmol), 1 μL of 10-fold diluted DNA template (approximately 1 ng), 2.5 μL of 10-fold PCR buffer, 1.5 μL of MgCl2 (50 mM ), and lastly...