iscript reverse transcription supermix kit Search Results


one step rt qpcr  (New England Biolabs)
99
New England Biolabs one step rt qpcr
One Step Rt Qpcr, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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strand cdna synthesis kit  (TaKaRa)
99
TaKaRa strand cdna synthesis kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Strand Cdna Synthesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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high-capacity cdna reverse transcription kit  (Thermo Fisher)
90
Thermo Fisher high-capacity cdna reverse transcription kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
High Capacity Cdna Reverse Transcription Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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stratascript rt-pcr kit  (Agilent technologies)
90
Agilent technologies stratascript rt-pcr kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Stratascript Rt Pcr Kit, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1st strand cdna synthesis kit  (Boehringer Mannheim)
90
Boehringer Mannheim 1st strand cdna synthesis kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
1st Strand Cdna Synthesis Kit, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 90 stars, based on 1 article reviews
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one-step rt-pcr kit  (Toyobo)
90
Toyobo one-step rt-pcr kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
One Step Rt Pcr Kit, supplied by Toyobo, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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amv reverse transcriptase kit  (Promega)
90
Promega amv reverse transcriptase kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Amv Reverse Transcriptase Kit, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hiscript ii q rt supermix for qpcr  (Vazyme Biotech Co)
96
Vazyme Biotech Co hiscript ii q rt supermix for qpcr
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Hiscript Ii Q Rt Supermix For Qpcr, supplied by Vazyme Biotech Co, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/hiscript ii q rt supermix for qpcr/product/Vazyme Biotech Co
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hiscript iii 1st strand cdna synthesis kit +gdna wiper  (Vazyme Biotech Co)
99
Vazyme Biotech Co hiscript iii 1st strand cdna synthesis kit +gdna wiper
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Hiscript Iii 1st Strand Cdna Synthesis Kit +Gdna Wiper, supplied by Vazyme Biotech Co, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/hiscript iii 1st strand cdna synthesis kit +gdna wiper/product/Vazyme Biotech Co
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ultrascript reverse transcriptase  (PCR Biosystems Ltd)
96
PCR Biosystems Ltd ultrascript reverse transcriptase
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Ultrascript Reverse Transcriptase, supplied by PCR Biosystems Ltd, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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artus® hcv rg rt-pcr kit  (Qiagen)
90
Qiagen artus® hcv rg rt-pcr kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Artus® Hcv Rg Rt Pcr Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/artus® hcv rg rt-pcr kit/product/Qiagen
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magmax viral rna isolation kit  (Thermo Fisher)
96
Thermo Fisher magmax viral rna isolation kit
Gamma-irradiated SARS-CoV-2 virus (BEI Resources) at 2 genome equivalents/uL was spiked in 1mL of saliva kept either in non-GTR STM (“Saliva”) (open circles) or GTR-STM devices (open diamonds), extracted with <t>MagMAX</t> Viral <t>RNA</t> Kit (ThermoFisher) and RT-PCR performed with CDC’s N1 primer. The pass/fail criteria set at 35.7 CT is 3 CT values more than the average CT value of the “Saliva” only samples. “Saliva” samples without GTR-STM gave a mean CT of 32.4 CT (Std Dev, ±0.3), and Saliva Samples in GTR-STM gave a mean CT of 32.7 CT (Std Dev, ±0.2). Study setup Experimental Sample : A contrived GTR-STM sample (n=9) was prepared by spiking 1mL of saliva with 2 genome equivalents/uL of -gamma-irradiated SARS-CoV-2 virus. Control sample : A contrived non-GTR STM (“Saliva”) (n=3) sample was prepared by spiking 1mL of saliva with 2 genome equivalents/uL of gamma-irradiated SARS-CoV-2 virus. Sample Extraction :RNA was extracted from 200uL of sample from both control and stressed samples following manufacturer’s instructions for MagMAX Viral RNA (ThermoFisher) manual protocol and eluted with 50uL of elution buffer. Quantification : Amplify 5uL of extracted RNA from each sample in triplicates with TaqPath master mix (ThermoFisher) and CDC’s N1 Primer (IDT).
Magmax Viral Rna Isolation Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Figure 6. Southern blot analysis of PCR products obtained from total RNA isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml cDNA mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.

Journal: Stem cells (Dayton, Ohio)

Article Title: Adenovirus mediated alpha interferon (IFN-alpha) gene transfer into CD34+ cells and CML mononuclear cells.

doi: 10.1002/stem.150386

Figure Lengend Snippet: Figure 6. Southern blot analysis of PCR products obtained from total RNA isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml cDNA mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.

Article Snippet: RNA is reverse-transcribed using the first strand cDNA synthesis kit from Clontech, Inc. (Palo Alto, CA).

Techniques: Southern Blot, Isolation, Transfection, Amplification, Control, Clone Assay, Electrophoresis

Figure 7. Southern blot analysis of PCR products obtained from total RNA isolated from CML hematopoietic stem cells (BMMNC) transfected with AdCMV-IFN-α gene. Lane 1, PCR products amplified from nontransfected (control) cell RNA; lane 2, RNA obtained from cells transfected with AdCMV-IFN-α after 24 h; lane 3, RNA from CFU-GM clones after 12 days; lane 4, CFU-GM clones after 14 days of culture; lane 5, no RNA was added; and lane 6, positive control. Total RNA was extracted from stem cells or CFU-GM clones by one single extraction. cDNA was synthesized from total RNA. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml cDNA mixture of a total reaction vol- ume of 25 ml. Aliquots of the reaction mixture (10 ml) were sub- jected to electrophoresis in a 0.8% agarose (Sigma) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, using α-32P-CTP. Filters were exposed to x-ray for 48 h at –80°C.

Journal: Stem cells (Dayton, Ohio)

Article Title: Adenovirus mediated alpha interferon (IFN-alpha) gene transfer into CD34+ cells and CML mononuclear cells.

doi: 10.1002/stem.150386

Figure Lengend Snippet: Figure 7. Southern blot analysis of PCR products obtained from total RNA isolated from CML hematopoietic stem cells (BMMNC) transfected with AdCMV-IFN-α gene. Lane 1, PCR products amplified from nontransfected (control) cell RNA; lane 2, RNA obtained from cells transfected with AdCMV-IFN-α after 24 h; lane 3, RNA from CFU-GM clones after 12 days; lane 4, CFU-GM clones after 14 days of culture; lane 5, no RNA was added; and lane 6, positive control. Total RNA was extracted from stem cells or CFU-GM clones by one single extraction. cDNA was synthesized from total RNA. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml cDNA mixture of a total reaction vol- ume of 25 ml. Aliquots of the reaction mixture (10 ml) were sub- jected to electrophoresis in a 0.8% agarose (Sigma) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, using α-32P-CTP. Filters were exposed to x-ray for 48 h at –80°C.

Article Snippet: RNA is reverse-transcribed using the first strand cDNA synthesis kit from Clontech, Inc. (Palo Alto, CA).

Techniques: Southern Blot, Isolation, Transfection, Amplification, Control, Clone Assay, Positive Control, Extraction, Synthesized, Electrophoresis

Gamma-irradiated SARS-CoV-2 virus (BEI Resources) at 2 genome equivalents/uL was spiked in 1mL of saliva kept either in non-GTR STM (“Saliva”) (open circles) or GTR-STM devices (open diamonds), extracted with MagMAX Viral RNA Kit (ThermoFisher) and RT-PCR performed with CDC’s N1 primer. The pass/fail criteria set at 35.7 CT is 3 CT values more than the average CT value of the “Saliva” only samples. “Saliva” samples without GTR-STM gave a mean CT of 32.4 CT (Std Dev, ±0.3), and Saliva Samples in GTR-STM gave a mean CT of 32.7 CT (Std Dev, ±0.2). Study setup Experimental Sample : A contrived GTR-STM sample (n=9) was prepared by spiking 1mL of saliva with 2 genome equivalents/uL of -gamma-irradiated SARS-CoV-2 virus. Control sample : A contrived non-GTR STM (“Saliva”) (n=3) sample was prepared by spiking 1mL of saliva with 2 genome equivalents/uL of gamma-irradiated SARS-CoV-2 virus. Sample Extraction :RNA was extracted from 200uL of sample from both control and stressed samples following manufacturer’s instructions for MagMAX Viral RNA (ThermoFisher) manual protocol and eluted with 50uL of elution buffer. Quantification : Amplify 5uL of extracted RNA from each sample in triplicates with TaqPath master mix (ThermoFisher) and CDC’s N1 Primer (IDT).

Journal: medRxiv

Article Title: COVID-19 Diagnostic Testing For All - Using Non-Dilutive Saliva Sample Collection, Stabilization and Ambient Transport Devices

doi: 10.1101/2021.01.20.20243782

Figure Lengend Snippet: Gamma-irradiated SARS-CoV-2 virus (BEI Resources) at 2 genome equivalents/uL was spiked in 1mL of saliva kept either in non-GTR STM (“Saliva”) (open circles) or GTR-STM devices (open diamonds), extracted with MagMAX Viral RNA Kit (ThermoFisher) and RT-PCR performed with CDC’s N1 primer. The pass/fail criteria set at 35.7 CT is 3 CT values more than the average CT value of the “Saliva” only samples. “Saliva” samples without GTR-STM gave a mean CT of 32.4 CT (Std Dev, ±0.3), and Saliva Samples in GTR-STM gave a mean CT of 32.7 CT (Std Dev, ±0.2). Study setup Experimental Sample : A contrived GTR-STM sample (n=9) was prepared by spiking 1mL of saliva with 2 genome equivalents/uL of -gamma-irradiated SARS-CoV-2 virus. Control sample : A contrived non-GTR STM (“Saliva”) (n=3) sample was prepared by spiking 1mL of saliva with 2 genome equivalents/uL of gamma-irradiated SARS-CoV-2 virus. Sample Extraction :RNA was extracted from 200uL of sample from both control and stressed samples following manufacturer’s instructions for MagMAX Viral RNA (ThermoFisher) manual protocol and eluted with 50uL of elution buffer. Quantification : Amplify 5uL of extracted RNA from each sample in triplicates with TaqPath master mix (ThermoFisher) and CDC’s N1 Primer (IDT).

Article Snippet: GTR-STM was tested by extraction with either QIAamp Viral RNA Mini Kit (Qiagen, cat #52906) or MagMAX Viral RNA Isolation Kit (ThermoFisher, cat #AM1939) and compared with either PBS or neat saliva as controls.

Techniques: Irradiation, Reverse Transcription Polymerase Chain Reaction

Heat-inactivated SARS-CoV-2 virus RNA (BEI Resources) at 500 genome equivalents/uL was spiked into 1mL of saliva kept either in GTR-STM collection devices or non-GTR-STM (“Saliva”) tubes, and stored at 25°C for 36 days. Matched spiked control saliva samples in both kinds of tubes were stored at -80°C. The pass/fail criteria are set at 32 CT value. 200uL of the sample is used at each time point to extract viral RNA with the MagMAX kit with a final elution volume of 50uL. The CT value of the viral RNA extracted with MagMAX viral RNA kit is normalized to input volume of 200uL (volume of sample used for RNA extraction). All GTR-STM samples gave excellent recoveries when compared to their matched -80°C control. Both the -80°C and the 25°C samples for the non-GTR-STM (“Saliva”) is above the pass/fail line even in the -80°C control samples indicating that the viral RNA is degraded by the Rnase in the short time (less than half hour) that the saliva sample is defrosting before RNA extraction is performed. Study setup Experimental Samples : 1 mL aliquots of saliva contrived with SARS-CoV-2 at 500 genome equivalents/uL (BEI Resources) were placed into either GTR-STM devices or non-GTR-STM (“Saliva”) tubes and stored at ambient (25°C) for up to 36 days. Control sample : 1 mL aliquots of saliva contrived with SARS-CoV-2 at 500 genome equivalents/uL (BEI Resources) were placed into either GTR-STM devices or non-GTR-STM (“Saliva”) tubes and stored at -80°C for up to 36 days. Sample Extraction : RNA extracted from 200uL of experimental and control samples with MagMAX viral RNA kit at days 10, 15, 20, 25, and 36 and eluted in 50uL of elution buffer. Quantification : 5uL of RNA was quantified with CDC’s SARS-CoV-2 RT-qPCR assay for N1 primer.

Journal: medRxiv

Article Title: COVID-19 Diagnostic Testing For All - Using Non-Dilutive Saliva Sample Collection, Stabilization and Ambient Transport Devices

doi: 10.1101/2021.01.20.20243782

Figure Lengend Snippet: Heat-inactivated SARS-CoV-2 virus RNA (BEI Resources) at 500 genome equivalents/uL was spiked into 1mL of saliva kept either in GTR-STM collection devices or non-GTR-STM (“Saliva”) tubes, and stored at 25°C for 36 days. Matched spiked control saliva samples in both kinds of tubes were stored at -80°C. The pass/fail criteria are set at 32 CT value. 200uL of the sample is used at each time point to extract viral RNA with the MagMAX kit with a final elution volume of 50uL. The CT value of the viral RNA extracted with MagMAX viral RNA kit is normalized to input volume of 200uL (volume of sample used for RNA extraction). All GTR-STM samples gave excellent recoveries when compared to their matched -80°C control. Both the -80°C and the 25°C samples for the non-GTR-STM (“Saliva”) is above the pass/fail line even in the -80°C control samples indicating that the viral RNA is degraded by the Rnase in the short time (less than half hour) that the saliva sample is defrosting before RNA extraction is performed. Study setup Experimental Samples : 1 mL aliquots of saliva contrived with SARS-CoV-2 at 500 genome equivalents/uL (BEI Resources) were placed into either GTR-STM devices or non-GTR-STM (“Saliva”) tubes and stored at ambient (25°C) for up to 36 days. Control sample : 1 mL aliquots of saliva contrived with SARS-CoV-2 at 500 genome equivalents/uL (BEI Resources) were placed into either GTR-STM devices or non-GTR-STM (“Saliva”) tubes and stored at -80°C for up to 36 days. Sample Extraction : RNA extracted from 200uL of experimental and control samples with MagMAX viral RNA kit at days 10, 15, 20, 25, and 36 and eluted in 50uL of elution buffer. Quantification : 5uL of RNA was quantified with CDC’s SARS-CoV-2 RT-qPCR assay for N1 primer.

Article Snippet: GTR-STM was tested by extraction with either QIAamp Viral RNA Mini Kit (Qiagen, cat #52906) or MagMAX Viral RNA Isolation Kit (ThermoFisher, cat #AM1939) and compared with either PBS or neat saliva as controls.

Techniques: RNA Extraction, Quantitative RT-PCR