rt buffer (Thermo Fisher)

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Thermo Fisher rt buffer
Rt Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rt+buffer/TRIS%2C+1%2E0M+buffer+soln%2E%2C+pH+8%2E4/pm42123321-240-56-85
Average 94 stars, based on 1 article reviews

rt buffer - by Bioz Stars, 2026-10

94/100 stars

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Reverse Transcription:

Article Title: Catch and release of sialoglycoRNAs enables sequencing-based profiling across cellular and extracellular material
Article Snippet: 20 μL of reverse transcription (RT) reaction was assembled by combining rPAL-labeled, sialidase-released glycoRNA with 0.5 μM anchored oligo-dT primer (5Bio-s7-dT15(VN), synthesized by IDT; sequence provided in Supplementary Table 1 ), 5% PEG-8000, and 0.5 mM dNTPs (NEB, cat. no. N0447), denatured at 72 °C for 3 min, and immediately cooled on ice. .. Reverse-transcription mix was then supplemented with 1× RT buffer (50 mM Tris-HCl, 75 mM KCl, 3 mM MgCl 2 , 10 mM DTT; pH 8.3), 1 mM GTP (NEB, cat. no. N0450), 40 U RiboLock RNase inhibitor (Thermo, cat. no. EO0381), 2 μM template-switching oligo (5Bio-s5-TSOr-N12; sequence provided in Supplementary Table 1 ), and 40 U Maxima H Minus RT enzyme (Thermo, cat. no. EP0752). .. Reaction was incubated at 42 °C for 90 min, followed by 10 cycles of 50 °C for 2 min and 42 °C for 2 min; then 85 °C for 5 min. RT product was amplified with 0.4 μM unique dual-indexed primer pairs (NEB, cat. no. E6440) in 100 μL PCR containing 1× SeqAmp PCR buffer with 2.5 U SeqAmp DNA polymerase (Takara, cat. no. 638509).

Article Title: Neocortical temporal patterning by a two-layered regulatory network
Article Snippet: Briefly, RNA was eluted into 4 μL elution mix containing 1 μL RT primer (10 μM), 1 μL dNTP mix (10 mM each, New England Biolabs, N0447L), 1 μL RNase Inhibitor (4 U/μL), and 1 μL H 2 O. Eluted samples were incubated at 72 °C for 3 min and immediately placed on ice. .. Each sample was added with 7 μL reverse transcription (RT) mix containing 0.75 μL H 2 O, 0.1 μL Maxima Reverse Transcriptase (Thermo Fisher Scientific, EP0741), 2 μL 5 × RT buffer (Thermo Fisher Scientific, EP0741), 2 μL Betaine (5 M, Sigma-Aldrich, B0300), 0.9 μL MgCl 2 (100 mM), 1 μL TSO primer (10 μM), 0.25 μL RNase Inhibitor (40 U/μL). ..

Article Title: Effect of Vitamin K Supplementation on Testosterone Production in a Rat Model of Late-Onset Hypogonadism
Article Snippet: .. The resulting RNA was denatured by incubation with 50 ng/μL oligo-dT primers (Hokkaido System Science Co., Sapporo, Japan) and 1 mM dNTP (GE Healthcare, Tokyo, Japan) at 65 ◦C for 5 min. Denatured RNA was used as a template to synthesize cDNA by mixing it with an RT buffer (50 mM Tris-HCl at pH 8.3, 75 mM KCl, 3 mM MgCl2, and 5 mM dithiothreitol) containing 50 U SuperScript III reverse transcriptase (Invitrogen, Carlsbad, CA, USA) and 20 U RNaseOUT RNase inhibitor (Invitrogen), followed by incubation at 50 ◦C for 60 min. .. The synthesized cDNA was used to amplify the target sequences using gene-specific primers (Table 1) and TB Green Premix Ex Taq (Takara Bio, Otsu, Japan) by qPCR in a CFX Connect Real-Time PCR Detection System https://doi.org/10.3390/foods15061070 (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Article Title: Proangiogenic effects of peritumoral adipose tissue in kidney cancer
Article Snippet: The RNA was first denatured at 70 °C for 5 min. in the presence of 2.5 μg of random hexamer primers (Thermo Fisher Scientific, Carlsbad, CA, USA). .. For the subsequent RT reaction, the following mixture was added: RT buffer [50 mM Tris–HCl (pH8.4), 75 mM KCl, 3 mM MgCl2], 0.5 mM dNTPs, 5 mM DTT, 200 units M-MLV Reverse Transcriptase (Thermo Fisher Scientific, Carlsbad, CA, USA). ..

Article Title: Effect of Vitamin K Supplementation on Testosterone Production in a Rat Model of Late-Onset Hypogonadism
Article Snippet: .. The resulting RNA was denatured by incubation with 50 ng/μL oligo-dT primers (Hokkaido System Science Co., Sapporo, Japan) and 1 mM dNTP (GE Healthcare, Tokyo, Japan) at 65 °C for 5 min. Denatured RNA was used as a template to synthesize cDNA by mixing it with an RT buffer (50 mM Tris-HCl at pH 8.3, 75 mM KCl, 3 mM MgCl 2 , and 5 mM dithiothreitol) containing 50 U SuperScript III reverse transcriptase (Invitrogen, Carlsbad, CA, USA) and 20 U RNaseOUT RNase inhibitor (Invitrogen), followed by incubation at 50 °C for 60 min. .. The synthesized cDNA was used to amplify the target sequences using gene-specific primers ( ) and TB Green Premix Ex Taq (Takara Bio, Otsu, Japan) by qPCR in a CFX Connect Real-Time PCR Detection System (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Article Title: Single-cell RNA-seq using UltraMarathonRT expands the known transcriptome
Article Snippet: .. For reverse transcription, 10 μL of RT buffer (100 mM Tris-HCl, pH 8.3 at 25°C (Invitrogen), 400 mM KCl (Ambion), 4 mM MgCl 2 (Ambion), 10 mM DTT (Sigma), 40 % glycerol (Promega)), 1 μL of uMRT (20 units/μL, RNAConnect), 0.5 μL of RNAseOUT (ThermoFisher), 1 μL Boost (if added, otherwise nuclease-free water, RNAConnect) and 1.5 μL nuclease-free water (Ambion) were added to a total volume of 20 μL. .. The reaction was incubated at 42°C for 60 min and inactivated at 95°C for 1 min. Next, 1 μL of 8 μM PCR_fwd01 primer (5’-GAAGTTAACAAGTAGAATCATTTATCCGGC-3’, IDT), 1 μL of 8 μM PCR_rev01primer (5’-TACGCTTACTCCAATACGACGCTC-3’, IDT), 10 μL LongAmp Taq 2X Master Mix (NEB), 7 μL nuclease-free water were added.

Sequencing:

Incubation:

Article Title: Effect of Vitamin K Supplementation on Testosterone Production in a Rat Model of Late-Onset Hypogonadism
Article Snippet: .. The resulting RNA was denatured by incubation with 50 ng/μL oligo-dT primers (Hokkaido System Science Co., Sapporo, Japan) and 1 mM dNTP (GE Healthcare, Tokyo, Japan) at 65 °C for 5 min. Denatured RNA was used as a template to synthesize cDNA by mixing it with an RT buffer (50 mM Tris-HCl at pH 8.3, 75 mM KCl, 3 mM MgCl 2 , and 5 mM dithiothreitol) containing 50 U SuperScript III reverse transcriptase (Invitrogen, Carlsbad, CA, USA) and 20 U RNaseOUT RNase inhibitor (Invitrogen), followed by incubation at 50 °C for 60 min. .. The synthesized cDNA was used to amplify the target sequences using gene-specific primers ( ) and TB Green Premix Ex Taq (Takara Bio, Otsu, Japan) by qPCR in a CFX Connect Real-Time PCR Detection System (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

Article Title: Identification of conserved RNA regulatory switches in living cells using RNA secondary structure ensemble mapping and covariation analysis.
Article Snippet: RNA molecules can populate ensembles of alternative structural conformations; however, comprehensively mapping RNA conformational landscapes within living cells presents notable challenges and has, as such, so far remained elusive.. Here, we generate transcriptome-scale maps of RNA secondary structure ensembles in both Escherichia coli and human cells, uncovering features of structurally heterogeneous regions.. By combining ensemble deconvolution and covariation analyses, we report the discovery of several bacterial RNA thermometers in the 5′ untranslated regions (UTRs) of the cspG, cspI, cpxP and lpxP mRNAs of Escherichia coli.

other:

Article Title: Extracellular Vesicle from Chlorella vulgaris Alleviates Hepatic Fibrosis in a Mouse Model of Metabolic Dysfunction-Associated Steatotic Liver Disease Through Modulation of Inflammatory Signaling.
Article Snippet: Images were acquired using an Olympus https://doi.org/10.3390/ijms27093735 IX81S1F-3 (Olympus, Tokyo, Japan) with a 10× objective lens.

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