Log In Sign up
The largest database of trusted experimental protocols

Magmax ai nd viral rna isolation kit

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in United States

The MagMAX AI/ND Viral RNA Isolation Kit is a lab equipment product designed for the extraction and purification of viral RNA from various sample types. It utilizes magnetic bead technology to efficiently capture and isolate viral genetic material.

Automatically generated - may contain errors

Lab products found in correlation

Agpath id one step rt pcr kit, by Thermo Fisher Scientific (6 mentions) Abi 7500 fast real time pcr system, by Thermo Fisher Scientific (6 mentions) Trizol ls reagent, by Thermo Fisher Scientific (5 mentions) Onetaq one step rt pcr kit, by New England Biolabs (1 mentions) Trizol ls, by Thermo Fisher Scientific (1 mentions) Rneasy mini kit, by Qiagen (1 mentions)

Close spelling variants of this product

RNA isolation kit (161 citations) MagMAX Viral RNA Isolation Kit (104 citations) MagMAX-96 AI/ND Viral RNA Isolation Kit (39 citations) MagMax AI/ND RNA isolation kit (2 citations) MagMax viral RNA kit (2 citations) Ambion MagMAX-96 AI/ND Viral RNA Isolation kit (2 citations)

7 protocols using magmax ai nd viral rna isolation kit

1

qRT-PCR Detection of Newcastle Disease Virus

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total RNA was extracted from swab medium and quantified as previously described [38 (link)], Briefly, RNA was extracted using Trizol LS reagent to inactivate the virus (Invitrogen, Carlsbad, CA, USA) and the MagMAX AI/ND Viral RNA Isolation Kit (Ambion, Austin, TX, USA). qRT-PCR targeting the NDV M gene was performed using previously described primers [38 (link)], the AgPath-ID one-step RT-PCR Kit (Ambion, Austin, TX, USA), and the ABI 7500 Fast Real-Time PCR system (Applied Biosystems, Waltham, MA, USA). A standard curve for each virus was established with RNA extracted and diluted from the same titrated stock of the viruses used to inoculate ECE. For virus quantification, a standard curve was established with RNA extracted and diluted from the same titrated stock of the virus used for inoculation. The qRT-PCR limit of detection for lentogenic NDV strains was between 101.5 and 102.3 EID50/mL equivalents. The detection limit for vNDV was 101.5 EID50/mL equivalents. Chickens with viral levels below the limit of detection were recorded as shedding just below the limit of detection.
Marcano V.C., Cardenas-Garcia S., Diel D.G., Antoniassi da Silva L.H., Gogal RM J.r., Miller P.J., Brown C.C., Butt S.L., Goraichuk I.V., Dimitrov K.M., Taylor T.L., Williams-Coplin D., Olivier T.L., Stanton J.B, & Afonso C.L. (2021). A Novel Recombinant Newcastle Disease Vaccine Improves Post- In Ovo Vaccination Survival with Sustained Protection against Virulent Challenge. Vaccines, 9(9), 953.
+ Open protocol
+ Expand
2

RT-PCR Detection of Viral RNA

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Viral RNA was extracted using the MagMAX AI/ND Viral RNA Isolation Kit (Ambion, Austin, TX, USA). For experimental chicken swab samples and challenge virus inoculum, one step RT-PCR was conducted with 20 μL of RNA template in a final reaction volume of 50 μL using OneTaq® One-Step RT-PCR Kit (NEB, Ipswich, MA, USA) with the primers Optil-F1, 5′-GTTACGCGCCAGCAAAAGCAGG-3′, Optil-F2, 5′-GTTACGCGCCAGCGAAAG CAGG-3′, and Optil-R1, 5′-GTTACGCGCCAGTAGAAACAAGG-3′ as described previously [4 (link)]. The PCR cycling was performed as follows: 95 °C for 2 min, 42 °C for 60 min, 94 °C for 2 min, 5 cycles of 94 °C for 30 s, 44 °C for 30 s, and 68 °C for 3.5 min, followed by 26 cycles of 94 °C for 30 s, 57 °C for 30 s, 68 °C for 3.5 min with a final extension at 68 °C for 10 min.
Chrzastek K., Segovia K., Torchetti M., Killian M.L., Pantin-Jackwood M, & Kapczynski D.R. (2021). Virus Adaptation Following Experimental Infection of Chickens with a Domestic Duck Low Pathogenic Avian Influenza Isolate from the 2017 USA H7N9 Outbreak Identifies Polymorphic Mutations in Multiple Gene Segments. Viruses, 13(6), 1166.
+ Open protocol
+ Expand
3

Rapid Detection of Avian Influenza and Newcastle Disease Viruses

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
OP and CL swabs were collected in 2 mL of BHI broth with a final concentration of gentamicin (1,000 μg/mL), penicillin G (10,000 units/mL), and amphotericin B (20 IU/mL) and kept frozen at −70°C until processed. Viral RNA was extracted using Trizol LS reagent (Invitrogen, Calsbad, CA) and the MagMAX AI/ND Viral RNA Isolation Kit (Ambion, Austin, TX, USA). Quantitative real time RT-PCR (qRT-PCR) for AIV and NDV detection was performed as previously described (Costa-Hurtado et al., 2014 (link)). qRT-PCR reactions targeting the influenza virus M gene (Spackman et al., 2002 (link)) and NDV M gene (Wise et al., 2004 (link)) were conducted using AgPath-ID one-step RT-PCR Kit (Ambion, Austin, TX, USA) and the ABI 7500 Fast Real-Time PCR system (Applied Biosystem, Calsbad, CA, USA). The RT step conditions for both primer sets were 10 min at 45°C and 95°C for 10 min. The cycling conditions for AIV were 45 cycles of 15 s, 95°C; 45 s, 60°C; and for NDV were 40 cycles of 10 s, 94°C; 30 s, 56°C; 10 s, 72°C. The calculated qRT-PCR lower detection limit for LPAIV, HPAIV and vNDV was 100.8EID50/mL, 101.0EID50/mL, and 100.94EID50/mL, respectively. A standard curve for virus quantification was established with RNA extracted from dilutions of the same titrated stock of the challenge virus, and results also reported as EID50/mL equivalents.
Pantin-Jackwood M., Costa-Hurtado M., Miller P.J., Afonso C.L., Spackman E., Kapczynski D., Shepherd E., Smith D, & Swayne D. (2015). Experimental co-infections of domestic ducks with a virulent Newcastle disease virus and low or highly pathogenic avian influenza viruses. Veterinary microbiology, 177(0), 7-17.
+ Open protocol
+ Expand
4

Viral RNA Extraction and Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols
Viral RNA was extracted using Trizol LS reagent (Invitrogen, Carlsbad, CA, USA) and the MagMAX AI/ND Viral RNA Isolation Kit (Ambion, Austin, TX, USA). Quantitative real-time RT-PCR (RRT-PCR) was performed using primers and a probe specific for the type A avian influenza (AI) matrix gene (Spackman, 2002 #104). The reactions were conducted using AgPath-ID one-step RT-PCR Kit (Ambion) and the ABI 7500 Fast Real-Time PCR system (Applied Biosystems, Carlsbad, CA, USA). For virus quantification, a standard curve was established with the RNA extracted from dilutions of the same titrated stock of the challenge virus. Cycle threshold (CT) values of each viral dilution were plotted against viral titers. The resulting standard curve had a high correlation coefficient (R2 > 0.99), and it was used to convert CT values to EID50/mL [2 (link),18 (link)]. The lower limit of detection was 0.9 × log10 EID50 per mL.
Chrzastek K., Segovia K., Torchetti M., Killian M.L., Pantin-Jackwood M, & Kapczynski D.R. (2021). Virus Adaptation Following Experimental Infection of Chickens with a Domestic Duck Low Pathogenic Avian Influenza Isolate from the 2017 USA H7N9 Outbreak Identifies Polymorphic Mutations in Multiple Gene Segments. Viruses, 13(6), 1166.
+ Open protocol
+ Expand
5

Quantification of Influenza Viral RNA

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Oropharyngeal and cloacal swabs were collected and kept frozen at −70°C. Viral RNA was extracted using Trizol LS (Invitrogen, Carlsbad, CA) and the MagMAX AI/ND Viral RNA Isolation Kit (Ambion, Austin, TX). Quantitative real time RT-PCR (qRRT-PCR) was performed as previously described (Kapczynski et al., 2016 (link)). Briefly, qRRT-PCR targeting the influenza M gene was conducted using AgPath-ID one-step RT-PCR Kit (Ambion) and the ABI 7500 Fast Real-Time PCR system (Applied Biosystems, Carlsbad, CA). For viral quantification, a standard curve was established with viral RNA extracted from the titrated challenge virus. Results were reported as EID50/ml equivalents and the lower limit of detection being 100.9EID50/ml for samples from chickens.
Pushko P., Tretyakova I., Hidajat R., Zsak A., Chrzastek K., Tumpey T.M, & Kapczynski D.R. (2016). Virus-like particles displaying H5, H7, H9 hemagglutinins and N1 neuraminidase elicit protective immunity to heterologous avian influenza viruses in chickens. Virology, 501, 176-182.
+ Open protocol
+ Expand
6

QRRT-PCR for Influenza Virus Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols
Oropharyngeal and cloacal swabs were collected in sterile brain heart infusion medium and kept frozen at −70°C. Viral RNA was extracted using Trizol LS reagent (Invitrogen, Calsbad, CA) and the MagMAX AI/ND Viral RNA Isolation Kit (Ambion, Austin, TX). Quantitative real time RT-PCR (qRRT-PCR) was performed as previously described (Pantin-jackwood, 2013). Briefly, qRRT-PCR targeting the influenza M gene was conducted using AgPath-ID one-step RT-PCR Kit (Ambion) and the ABI 7500 Fast Real-Time PCR system (Applied Biosystems, Calsbad, CA). For viral quantification, a standard curve was established with viral RNA extracted from the titrated challenge virus, GyrF H5N8, Ck/WJ H5N1 or Ck/Egypt H5N1. Results were reported as EID50/ml equivalents and the lower limit of detection being 100.9 EID50/ml for samples from chickens.
Kapczynski D.R., Tumpey T.M., Hidajat R., Zsak A., Chrzastek K., Tretyakova I, & Pushko P. (2016). Vaccination with virus-like particles containing H5 antigens from three H5N1 clades protects chickens from H5N1 and H5N8 influenza viruses. Vaccine, 34(13), 1575-1581.
+ Open protocol
+ Expand
7

Quantifying Avian Influenza Viral RNA

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Viral RNA was extracted from swabs using the MagMAX AI/ND Viral RNA Isolation Kit (Ambion, Austin, TX, USA). Quantitative real time RT-PCR (qRRT-PCR) for AIV detection was performed as previously described (20 (link)). qRRT-PCR reactions targeting the influenza virus M gene (25 (link)) were conducted using AgPath-ID one-step RT-PCR Kit (Ambion, Austin, TX, USA) and the ABI 7500 Fast Real-Time PCR system (Applied Biosystems, Carlsbad, CA, USA). The RT step conditions were 10 min at 45°C and 95°C for 10 min. The cycling conditions were 45 cycles of 15 s, 95°C; 45 s, 60°C. Virus titers in frozen tissue samples were determined by weighing, homogenizing, and diluting tissues in BHI to a 10% (wt/vol) concentration. Viral RNA was extracted using Trizol LS reagent (Invitrogen, Carlsbad, CA) and the Qiagen RNeasy Mini Kit (Qiagen, USA). Equal amounts of RNA extracted from the tissue samples were used in the qRRT-PCR assay (50 ng/μl). For virus quantification, a standard curve was established with RNA extracted from dilutions of the same titrated stock of the challenge virus, and results reported as EID50/ml or EID50/gr equivalents. The calculated qRT-PCR lower detection limit for the viruses varied between 101.5EID50/ml, and 102.5EID50/ml.
DeJesus E., Costa-Hurtado M., Smith D., Lee D.H., Spackman E., Kapczynski D.R., Torchetti M.K., Killian M.L., Suarez D.L., Swayne D.E, & Pantin-Jackwood M.J. (2016). Changes in adaptation of H5N2 highly pathogenic avian influenza H5 clade 2.3.4.4 viruses in chickens and mallards. Virology, 499, 52-64.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!