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Luna universal probe one step rt pcr kit

Manufactured by New England Biolabs
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The Luna Universal Probe One-Step RT-PCR Kit is a reagent system for performing one-step reverse transcription and real-time PCR amplification of RNA targets using fluorescent probe detection.

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Lab products found in correlation

Purelink rna mini kit, by Thermo Fisher Scientific (2 mentions) Bead miller, by Thermo Fisher Scientific (2 mentions) Abi 7500 fast dx, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Luna Universal One-Step RT-PCR kit (9 citations) RT-PCR kit (3 citations) Luna Universal Probe one-step reverse transcription-quantitative PCR (RT-qPCR) kit (2 citations)

3 protocols using luna universal probe one step rt pcr kit

1

Quantifying mRNA Vaccine RNA Levels

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Tissues were taken at day 1 post-immunization. Tissues were weighted and homogenized with a bead miller (Fisher). Total RNA was purified with a PureLink RNA mini kit (Invitrogen). RNA purified from BNT162b2 mRNA vaccine was diluted serially and served as the standard. A Luna universal probe one-step RT-PCR kit (NEB) was used to measure cycle threshold (Ct) of purified RNA with mRNA vaccine-specific primers (mVac-F: 5’-TACCAAGCTGAACGACCTGT, mVac-R: 5’-TTGCTGTTCCAGGCAATCAC, mVac-Probe: 5’-FAM-TGCCCGACGACTTCACCGGC-IBFQ).
Li C., Lee A., Grigoryan L., Arunachalam P.S., Scott M.K., Trisal M., Wimmers F., Sanyal M., Weidenbacher P.A., Feng Y., Adamska J.Z., Valore E., Wang Y., Verma R., Reis N., Dunham D., O’Hara R., Park H., Luo W., Gitlin A.D., Kim P., Khatri P., Nadeau K.C, & Pulendran B. (2022). Mechanisms of innate and adaptive immunity to the Pfizer-BioNTech BNT162b2 vaccine. Nature immunology, 23(4), 543-555.
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2

Quantifying mRNA Vaccine RNA Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tissues were taken at day 1 post-immunization. Tissues were weighted and homogenized with a bead miller (Fisher). Total RNA was purified with a PureLink RNA mini kit (Invitrogen). RNA purified from BNT162b2 mRNA vaccine was diluted serially and served as the standard. A Luna universal probe one-step RT-PCR kit (NEB) was used to measure cycle threshold (Ct) of purified RNA with mRNA vaccine-specific primers (mVac-F: 5’-TACCAAGCTGAACGACCTGT, mVac-R: 5’-TTGCTGTTCCAGGCAATCAC, mVac-Probe: 5’-FAM-TGCCCGACGACTTCACCGGC-IBFQ).
Li C., Lee A., Grigoryan L., Arunachalam P.S., Scott M.K., Trisal M., Wimmers F., Sanyal M., Weidenbacher P.A., Feng Y., Adamska J.Z., Valore E., Wang Y., Verma R., Reis N., Dunham D., O’Hara R., Park H., Luo W., Gitlin A.D., Kim P., Khatri P., Nadeau K.C, & Pulendran B. (2022). Mechanisms of innate and adaptive immunity to the Pfizer-BioNTech BNT162b2 vaccine. Nature immunology, 23(4), 543-555.
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3

SARS-CoV-2 Detection using RT-PCR

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The RT-PCR assay was performed using the ABI 7500 Fast DX (Applied Biosystems) instrument. We performed assays for the E and RNase P genes separately in 20-µL reaction volumes using the Luna Universal Probe One-Step RT-PCR Kit (New England Biolabs). The final concentrations of primer and probe were 400 and 200 nM, respectively. We followed the recommended protocols in Corman et al. (1 ) and the Stanford clinical virology laboratory’s SARS-CoV-2 RT-PCR test (14 ). For quantification from clinical samples, we used 8 and 2 µL of ITP-extracted nucleic acids for the E gene and RNase P gene reactions, respectively. For the E gene standard curve, we used 5 µL of various dilutions of synthetic RNA controls as template.
Ramachandran A., Huyke D.A., Sharma E., Sahoo M.K., Huang C., Banaei N., Pinsky B.A, & Santiago J.G. (2020). Electric field-driven microfluidics for rapid CRISPR-based diagnostics and its application to detection of SARS-CoV-2. Proceedings of the National Academy of Sciences of the United States of America, 117(47), 29518-29525.
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