Journal: Viruses

Article Title: Processing Hundreds of SARS-CoV-2 Samples with an In-House PCR-Based Method without Robotics

doi: 10.3390/v13091712

Figure Lengend Snippet: CFX plate loading scheme. Shown is the pipetting strategy for the transfer of extracted RNA from the 96-well elution plate to the 384-well qPCR plate using a 12-channel pipette in order to run a full plate RT-qPCR experiment that includes 88 samples measured with three primer/probe sets like N1 (blue), N2 (orange) and RP (purple). Four dilutions (10 5 , 10 4 , 10 3 , and 10 2 copies) of the RNA control as well as two no-template control reactions (NTC) for each primer/probe set are included.

Article Snippet: The RT-qPCR reactions were set up in an optical 384-well plate (Bio-Rad, Hercules, CA, USA, #HSP3865) on ice and sealed with adhesive optical clear qPCR plate seals (Bio-Rad, #MSB1001), briefly mixed and centrifuged for 2 min at 2000× g . The RT-qPCR run was performed using a CFX384 Real-Time System type C1000 thermocycler (Bio-Rad) with the following thermal conditions: 25 °C for 2 min, 50 °C for 20 min, 95 °C for 3 min and 45 cycles of 95 °C for 15 s and 58 °C for 10 s including a plate read to detect for the FAM/Cy5/HEX signal, depending on the probes used.

Techniques: Transferring, Quantitative RT-PCR, Control