Figure 1. Stability of RNA and DNA stored in Norgen's Total Nucleic Acid Preservative. GAPDH (for DNA) and β-actin (for RNA) were monitored over an 18 week period for DNA and 60 days for RNA from swab samples stored at ambient temperature in Norgen’s Total Nucleic Acid Preservation Tubes. RNA was isolated from 250 µL of preserved swab samples on day 0, day 4, day 6, day 10, day 16, day 30 and day 60 using Norgen’s Total RNA Isolation Kit (Cat.# 17200). DNA was isolated from 250 µL of preserved swab samples on day 0 and 18 weeks using Norgen’s Saliva DNA Isolation Kit (Cat# Dx45400). Next, 4 µL of elution was used in a 20 µL real-time PCR reaction using CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad). RNA can be stored in Norgen’s Total Nucleic Acid Preservative for up to 60 days and DNA can be stored for up to 18 weeks at room temperature with no change in quality and quantity of RNA and DNA.
|Cat. Dx69200||Total Nucleic Acid Preservation Tubes Dx (For Diagnostic Use)||50 units||View product page|
|Cat. 69200||Total Nucleic Acid Preservation Tubes (For Research Use Only)||50 units||View product page|
|Cat. 53800||Saliva RNA Collection and Preservation Devices Dx (For Diagnostic Use)||50 devices||View product page|
|Cat. RU53800||Saliva RNA Collection and Preservation Devices (For Research Use Only)||50 devices||View product page|
Figure 1. SARS-CoV-2 E-gene and RdRP Amplification from Preserved Swabs and Saliva RNA isolated by Norgen’s Saliva/Swab RNA Purification Kit (Cat. #69100). Duplicate Nasopharyngeal swabs, Oropharyngeal Swabs and Saliva samples were collected from different donors. Nasopharyngeal and Oropharyngeal swab samples were collected into Norgen’s Total Nucleic Acid Preservative Tubes (Cat. 69200). Saliva samples were collected into Norgen’s Saliva RNA Collection and Preservation Devices (Cat. RU53800). All collected samples were spiked with 3 different concentrations from Norgen’s E/RdRP/RP Positive Control (included in Norgen’s COVID-19 TaqMan RT-PCR Kit (E/RdRP genes – TM67200) and processed for RNA isolation using Norgen’s Saliva/Swab RNA Purification Kit (Cat. #69100), using an input of 0.25 mL from each preserved samples. The sensitivity of the isolation method was assessed amplifying the spiked SARS-CoV-2 spiked targets using Norgen’s COVID-19 TaqMan RT-PCR Kit (E/RdRP genes (TM67200). Figure 1 shows the linear recovery of the different spike-in amounts indicating the high sensitivity of the isolation procedure as well as the high quality of the RNA purified using Norgen’s Saliva/Swab RNA Purification Kit (Cat. #69100).
|Dx69100||Saliva / Swab RNA Purification Kit Dx||50 Preps||View product page|
|69100||Saliva / Swab RNA Purification Kit||50 Preps||View product page|
|Cat. Dx69300||Saliva/Swab RNA Purification 96-Well Kit Dx||2 x 96-well plates||View product page|
|Cat. 69300||Saliva/Swab RNA Purification 96-Well Kit||2 x 96-well plates||View product page|
|Dx17200||Total RNA Purification Kit Dx||50 Preps||View product page|
|Total RNA Purification Kit||50 Preps,
|View product page|
|Total RNA Purification 96-Well Kits||2 x 96-well plates,
6 x 96-well plates
|View product page|
Background: Curative, Inc. developed an RT-PCR test for SARS-CoV-2 that makes use of Norgen’s Total RNA Purification 96-well Kit, #24350/#24380. They employ the use of this kit both manually or automated on the Tecan Resolvex A20. They have recently received received Emergency Use Authorization from the FDA for in vitro diagnostic use of this kit at KorvaLabs LLC.
"...RNA Extraction for all specimen types is performed using Total RNA Purification 96- Well Kit (Norgen Biotek Corporation) manually or using Tecan Resolvex A200. The input sample volume is 300µl, the elution volume is 60µl." (p.2)
Proceed to PCR Based Detection
Figure 1. Duplex TaqMan RT-PCR analytical sensitivity for each of the SARS-CoV-2 targets based on the Charité/Berlin protocol. The analytical sensitivity was determined by analyzing six serial dilutions (200,000 copies to 2 copies) of Norgen's positive control from the COVID-19 TaqMan RT-PCR Kit (E/RdRP gene)(Cat. TM67200). Five µL of template was used in a 20 µL of the one-step RT-PCR reaction and the arrow indicates a copy number of 10.
|Cat. TM67101||50 Rxns|
|Cat. TM67130||500 Rxns|
|Cat. 28113||100 Rxns|
|Cat. 28114||200 Rxns|
|Cat. 28115||500 Rxns|
Figure 1. Performance of Norgens 2X One-Step RT-PCR Master Mix (28113) to detect SARS-CoV-2 using Norgens 2019-nCoV Primer & Probe Mixes (TM67101) and Norgens 2019-nCoV RT-PCR Positive Control (PC67102). The one-step RT-PCR reaction was prepared according to the CDC protocol and the sensitivity of detection of the three targets; 2019-nCoV_ N1 (Circle), 2019-nCoV_ N2 (Triangle) and RNase P (Cross), was compared with Competitor Ns product (Red).
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