The spike-in control, including 3 distinct lambda DNA amplification products (~180 bp; one without modification and the other two with 5mC and 5hmC modifications), was prepared as follow47: lambda DNA was PCR amplified by Taq DNA Polymerase (NEB) and purified by AMPure XP beads (Beckman...
Real Time PCR analysis for viral DNA The amount of adenoviral genomes per cell was determined by a multiplex real-time polymerase chain reaction =-=[40]-=-. Real-time PCR is based on the 5'-3' nuclease activity of AmpliTaq Gold polymerase, which allows it to cleave fluorogenic probes ...
(NH4)2SO4, 3 mmol l − 1 MgCl2, 50 μmol l − 1 each dNTP, 480 nmol l − 1 of each specific primer for SEA deletion, 900 nmol l − 1 of each primer for SNP (rs3760053), 1 μmol l − 1 SYBR green and 0.02 units Taq DNA polymerase (Biolabs Co., Ltd., Ipswich,...
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 ...
PCR with variant-specific primers (Integrated DNA Technologies) using the Taq DNA-Polymerase (Thermo Fisher Scientific, Waltham, MA, USA) or the OneTaq DNA Polymerase (New England Biolabs) with GC-Rich Buffer was performed for breakpoint confirmation. Primer sequences are available upon request....
The specificity of each selected primer pair was observed via standard PCR on synthesized cDNA using the BIOTAQ DNA polymerase (Bioline, London, UK), and each amplification product was verified by 1.5% agarose gel electrophoresis. Table 1 RPKM (Reads Per Kilobase per Million mapped reads) ...
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...
Nuclear mitochondrial DNA ONT: Oxford Nanopore Technologies PCR: Polymerase chain reaction PERMANOVA: Permutational multivariate analysis of variance SFB: Short fragment buffer SUP: Super accurate WoRMS: World Register of Marine Species del Socorro Toxqui Rodríguez M, Naya-Català F, Sitjà-Bobadilla A...
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,...
PCR-based validation Presence or absence of inserts was checked by polymerase chain reaction (PCR) amplifications using the 2X Power Taq PCR mix (Bioteke Corp.) according to manufacturer's instructions and using primers designed within the Tnt1 sequence (5′ and 3′ sides of the Tnt1 ...