Saliva/Swab RNA Purification Kits Dx
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The purified RNA is intended for in vitro diagnostic use for medical purposes.
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Saliva/Swab RNA Purification Kits Dx
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Supporting Data
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 sample. The sensitivity of the isolation method was assessed by 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).
Figure 2. RNase P Gene Amplification of Fresh and Preserved Swab and Saliva RNA isolated by Norgen’s Saliva/Swab RNA Purification Kit (Cat. #Dx69100). Duplicate preserved (P) and non-preserved (NP) Nasopharyngeal swabs, Oropharyngeal Swabs, and Saliva samples were collected from different donors. Preserved Nasopharyngeal and Oropharyngeal swab samples were collected into Norgen’s Total Nucleic Acid Preservative Tubes. Preserved Saliva samples were collected into Norgen’s Saliva RNA Collection and Preservation Devices. Non-preserved samples were processed immediately without mixing with any preservative. All collected samples were processed for RNA isolation using Norgen’s Saliva/Swab RNA Purification Kit (Cat. #Dx69100), using an input of 0.25 mL from preserved samples or the entire non-preserved swab for the non-preserved conditions. Non-preserved saliva samples were isolated from a 0.25mL input as well. Isolated RNA was tested by amplification of the RNase P gene by RT-PCR.
Figure 3. Human miR-21 Amplification of Fresh and Preserved Swab and Saliva RNA isolated by Norgen’s Saliva/Swab RNA Purification Kit (Cat. #Dx69100). Duplicate preserved (P) and non-preserved (NP) Nasopharyngeal swabs, Oropharyngeal Swabs, and Saliva samples were collected from different donors. Preserved Nasopharyngeal and Oropharyngeal swab samples were collected into Norgen’s Total Nucleic Acid Preservative Tubes. Preserved Saliva samples were collected into Norgen’s Saliva RNA Collection and Preservation Devices. Non-preserved samples were processed immediately without mixing with any preservative. All collected samples were processed for RNA isolation using Norgen’s Saliva/Swab RNA Purification Kit (Cat. #Dx69100), using an input of 0.25 mL from preserved samples or the entire non-preserved swab for the non-preserved conditions. Non-preserved saliva samples were isolated from a 0.25mL input as well. Isolated RNA was tested by amplification of the human miR-21 detected by stem-loop RT-PCR.
Figure 4. Average RNA Yield of Samples Isolated by Norgen’s Saliva/Swab RNA Purification Kit (Cat. #Dx69100). The average yield of RNA isolated by Norgen’s Saliva/Swab RNA Purification Kit (Cat. #Dx69100) is ≥ 1 µg. Duplicate preserved (P) and non-preserved (NP) swab (nasal and oral) and saliva samples were collected from different donors. Preserved swab samples (nasal or oral) were collected into Norgen’s Total Nucleic Acid Preservative Tubes (Lot #598129) and preserved saliva samples were collected using Norgen’s Saliva RNA Collection and Preservation Devices. Non-preserved samples were processed immediately without mixing with any preservative. All collected samples were processed for RNA isolation using Norgen’s Saliva/Swab RNA Purification Kit, using an input of 0.25 mL from preserved samples and the non-preserved saliva or the entire non-preserved swab for the non-preserved swabs. NanoDrop was used to estimate the concentration and yield of the isolated RNA.
Figure 5. SARS-CoV-2 E-gene and RdRP Amplification from Preserved Swabs and Saliva RNA isolated by Norgen’s Saliva/Swab RNA Purification 96-Well Kit (Cat. Dx69300). 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. Dx69200). Saliva samples were collected into Norgen’s Saliva RNA Collection and Preservation Devices (Cat. 53800). 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 96-Well Kit (Cat. Dx69300), using an input of 0.25 mL from each preserved sample. The sensitivity of the isolation method was assessed by amplifying the spiked SARS-CoV-2 spiked targets using Norgen’s COVID-19 TaqMan RT-PCR Kit (E/RdRP genes (DxTM67200). Figure 5 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 96-Well Kit (Cat. Dx69300).
Figure 6. RNase P Gene Amplification of Fresh and Preserved Swab and Saliva RNA isolated by Norgen’s Saliva/Swab RNA Purification 96-Well Kit Dx (Cat. #Dx69300). Duplicate preserved (P) and non-preserved (NP) Nasopharyngeal swabs, Oropharyngeal Swabs, and Saliva samples were collected from different donors. Preserved Nasopharyngeal and Oropharyngeal swab samples were collected into Norgen’s Total Nucleic Acid Preservative Tubes Dx (Cat # Dx69300). Preserved Saliva samples were collected into Norgen’s Saliva RNA Collection and Preservation Devices Dx (53800). Non-preserved samples were processed immediately without mixing with any preservative. All collected samples were processed for RNA isolation using Norgen’s Saliva/Swab RNA Purification 96-Well Kit Dx, using an input of 0.25 mL from preserved samples or the entire non-preserved swab for the non-preserved conditions. Non-preserved saliva samples were isolated from a 0.25mL input as well. Isolated RNA was tested by amplification of the RNase P gene by RT-PCR.
Figure 7. Human miR-21 Amplification of Fresh and Preserved Swab and Saliva RNA isolated by Norgen’s Saliva/Swab RNA Purification 96-Well Kit Dx (Cat. #Dx69300). Duplicate preserved (P) and non-preserved (NP) Nasopharyngeal swabs, Oropharyngeal Swabs, and Saliva samples were collected from different donors. Preserved Nasopharyngeal and Oropharyngeal swab samples were collected into Norgen’s Total Nucleic Acid Preservative Tubes Dx (Cat. #Dx69200). Preserved Saliva samples were collected into Norgen’s Saliva RNA Collection and Preservation Devices Dx (Cat # 53800). Non-preserved samples were processed immediately without mixing with any preservative. All collected samples were processed for RNA isolation using Norgen’s Saliva/Swab RNA Purification 96-Well Kit Dx, using an input of 0.25 mL from preserved samples or the entire non-preserved swab for the non-preserved conditions. Non-preserved saliva samples were isolated from a 0.25mL input as well. Isolated RNA was tested by amplification of the human miR-21 detected by stem-loop RT-PCR.
Figure 8. Average RNA Yield of Samples Isolated by Norgen’s Saliva/Swab RNA Purification 96-Well Kit Dx (Cat. #Dx69300). Duplicate preserved (P) and non-preserved (NP) swab (nasal and oral) and saliva samples were collected from different donors. Preserved swab samples (nasal or oral) were collected into Norgen’s Total Nucleic Acid Preservative Tubes Dx and preserved saliva samples were collected using Norgen’s Saliva RNA Collection and Preservation Devices (Cat. #53800). Non-preserved samples were processed immediately without mixing with any preservative. All collected samples were processed for RNA isolation using Norgen’s Saliva/Swab RNA Purification 96-Well Kit Dx, using an input of 0.25 mL from preserved samples and the non-preserved saliva or the entire non-preserved swab for the non-preserved swabs. NanoDrop was used to estimate the concentration and yield of the isolated RNA.
Kit Specifications |
|
Sample Volume Range |
250 μL |
Size of RNA Purified |
All sizes, including small RNA (<200 nt) |
Minimum Elution Volume | 50 μL |
Maximum Elution Volume | 100 μL |
Time to Complete 10 Purifications | 15 - 20 minutes |
Average Yield |
≥ 1 μg * *Varies from sample to sample |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
Component | Cat. Dx69100 (50 preps) | Cat. Dx69300 (192 preps) |
---|---|---|
Lysis Buffer A | 30 mL | 100 mL |
Solution WN | 18 mL | 55 mL |
Wash Solution A | 18 mL | 2 x 38 mL |
Elution Solution A | 6 mL | 20 mL |
Mini Spin Columns | 50 | - |
96-Well Isolation Plate (Deep Well) | - | 2 |
Collection Tubes | 50 | - |
96-Well Collection Plate (Deep Well) | - | 2 |
Elution Tubes (1.7 mL) | 50 | - |
96-Well Elution Plate (Deep Well) | - | 2 |
Adhesive Tape | - | 4 |
Product Insert | 1 | 1 |
Automation
Supplementary Protocol
Supplementary Protocol - Automated Procedure for Saliva/Swab RNA Purification and Concentration
Application Note