PCR reactions with genomic DNA as a template were carried out in a total volume of 50 μl that contained ~100 ng of genomic DNA, 300 nM of each primer, 0.2 mM dNTP, 1.5 mM MgCl2, and 2.6 units of Expand High Fidelity enzyme mix in 1× Expand High Fidelity buffer (Roche Applied Science, Basel, Switzerland). The PCR reactions were performed on a T3 Thermocycler PCR system (Biometra, Göttingen, Germany) using the following conditions: 40 cycles of 94°C for 20 s, 55°C for 30 s, and 72°C for 45 s and a final extension was at 72°C for 10 min. The PCR products were cloned and sequenced (Eurofins, Ebersberg, Germany).
T3 thermocycler pcr system
Manufactured by Analytik Jena
Sourced in Germany, Switzerland
The T3 Thermocycler PCR system is a laboratory instrument designed for performing polymerase chain reaction (PCR) experiments. The core function of the T3 Thermocycler is to precisely control the temperature cycles required for DNA amplification during the PCR process.
Automatically generated - may contain errors
2 protocols using t3 thermocycler pcr system
1
High-Fidelity Genomic DNA Amplification
2
First-Strand cDNA Synthesis and RT-PCR
Check if the same lab product or an alternative is used in the 5 most similar protocols
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First strand cDNA synthesis was carried out using 1 μg of total RNA and Moloney murine leukemia virus (MMLV) reverse transcriptase lacking ribonuclease H activity (H minus) according to the manufacturer’s instructions (Promega). The subsequent PCR step was performed using the Expand High Fidelity PCR system (Roche Applied Science, Basel, Switzerland), a T3 Thermocycler PCR system (Biometra), and the following cycling conditions: 40 cycles of 94°C for 20 s, 55°C for 30 s, and 72°C for 45 s and a final extension at 72°C for 10 min. The RT-PCR products were cloned and sequenced.
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