NK-92 total RNA was extracted using the RNeasy Mini Kit (QIAGEN) according to the manufacturer’s protocol. Reverse transcription was performed using iScript Reverse Transcriptase and iScript Reaction mix per iScript cDNA Synthesis Kit manufacturer’s protocol (Bio-Rad). Real-Time PCR reactions were carried out using a PrimeTime Gene Expression Master Mix and custom PrimeTime Std qPCR assay primer/probe sets (IDT Technologies), as well as a TaqMan Gene Expression Assay (ThermoFisher). An ABI 7500 Real-Time PCR Thermal Cycler (ThermoFisher) was used under the PCR polymerase activation and amplification conditions of 95°C for 3 minutes and 40 cycles (95°C for 15 s and 60°C for 1 minute).
Abi 7500 real time pcr thermal cycler
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ABI 7500 Real-Time PCR Thermal Cycler is a laboratory instrument used for real-time polymerase chain reaction (PCR) analysis. It is designed to amplify and simultaneously quantify targeted DNA sequences. The ABI 7500 provides precise temperature control and data collection capabilities for real-time PCR applications.
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2 protocols using abi 7500 real time pcr thermal cycler
1
Real-Time PCR Analysis of NK-92 Cells
2
Multiplex PCR Panel for Viral Pathogens
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The one‐step RT‐PCR was fulfilled with the ABI Verity 96 Thermal Cycler. The PCR products were added to a 96‐well plate, prepared for capillary electrophoresis (CE), and fragment analysis by applying the 3500 Genetic Analyzer (ABI, USA), according to the manufacturer's protocol. The multiplex PCR panel included Mycoplasma pneumoniae and 8 viruses: enterovirus (EV), varicella‐zoster virus (VZV), mumps virus (MuV), cytomegalovirus (CMV), herpes simplex virus type 1 (HSV‐1), herpes simplex virus type 2 (HSV‐2), Epstein‐Barr virus (EBV), and human herpesvirus type 6 (HHV6).
The RT‐PCR was used to detect EV in CSF samples. The ABI 7500 real‐time PCR thermal cycler (Thermo Fisher, USA) and Real‐time TaqMan PCR reagents (Da'an gene Tech, China) were used to amplify the five targets separately according to the manufacturer's instructions.
The RT‐PCR was used to detect EV in CSF samples. The ABI 7500 real‐time PCR thermal cycler (Thermo Fisher, USA) and Real‐time TaqMan PCR reagents (Da'an gene Tech, China) were used to amplify the five targets separately according to the manufacturer's instructions.
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