Qscript cdna mix

Manufactured by Quantabio

Sourced in United States

QScript cDNA Mix is a reagent used for the reverse transcription of RNA into cDNA. It contains the necessary components for the conversion of RNA to cDNA, including a reverse transcriptase enzyme, buffer, and dNTPs.

Automatically generated - may contain errors

Lab products found in correlation

Sybr fast qpcr master mix, by Roche (4 mentions) Rneasy micro kit, by Qiagen (2 mentions) Rneasy mini kit, by Qiagen (1 mentions) Cfx connect real time pcr detection system, by Bio-Rad (1 mentions) Cfx manager 3, by Bio-Rad (1 mentions) Tri reagent, by Merck Group (1 mentions) Anti cd11b microbead, by Miltenyi Biotec (1 mentions) Anti acsa 2 microbead, by Miltenyi Biotec (1 mentions) Fc block, by Miltenyi Biotec (1 mentions) Papain dissociation system, by Worthington (1 mentions) Myelin removal beads 2, by Miltenyi Biotec (1 mentions)

Spelling variants of this product

QScript cDNA master mix (2 citations)

5 protocols using qscript cdna mix

Quantification of Akt Gene Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

RNA was isolated using the RNeasy Mini Kit (Qiagen Inc, Toronto, ON) according to manufacturer protocol. RNA was reverse transcribed using qScript cDNA mix from Quantabio (Beverly, MA). Gene expression was analyzed by qPCR reactions with SYBR Green qPCR Mastermix (Bioline Reagents Limited, London, ON) and performed on the CFX Connect Real-time PCR Detection system (Bio-Rad Canada, Mississauga, ON). Primers for human Akt1, Akt2, Akt3 and Hprt were purchased from Bio-Rad Canada (Mississauga, ON). Relative quantification was determined by normalizing expression to Hprt using CFX-Manager 3.1 (Bio-Rad Canada, Mississauga, ON).

Chorner P.M, & Moorehead R.A. (2018). A-674563, a putative AKT1 inhibitor that also suppresses CDK2 activity, inhibits human NSCLC cell growth more effectively than the pan-AKT inhibitor, MK-2206. PLoS ONE, 13(2), e0193344.

+ Open protocol

+ Expand

Astrocyte Isolation and RT-qPCR Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Astrocytes were isolated as previously described (52 (link)) with a few modifications. Briefly, mouse SC were minced into approximately 1 mm² pieces and digested using the Papain Dissociation System (Worthington Biochemical Corporation, catalog LK003153). Isolated cells were negatively selected in tandem first with Myelin Removal Beads II (Miltenyi Biotec, catalog 130-096-731) and second with anti-CD11b MicroBeads (Miltenyi Biotec, catalog 130-049-601) to collect the flowthrough fraction. To enrich astrocytes, the flowthrough cells were first treated with FcBlock (Miltenyi Biotec, catalog 130-092-575), then positively selected using anti-ACSA-2 MicroBeads (Miltenyi Biotec, catalog 130-097-678). Total RNA was prepared using TRI Reagent (MilliporeSigma, catalog 93289) and reverse-transcribed using qScript cDNA Mix (Quantabio, catalog 950048) to obtain cDNA. RT-qPCR assays were performed with SYBR FAST qPCR Master Mix (Kapa Biosystems, catalog KK4602), using primers indicated in Supplemental Table 2. Expression levels of target genes relative to an internal control, Actb, were calculated using the ΔΔCt method (53 (link)).

Barclay W.E., Aggarwal N., Deerhake M.E., Inoue M., Nonaka T., Nozaki K., Luzum N.A., Miao E.A, & Shinohara M.L. (2022). The AIM2 inflammasome is activated in astrocytes during the late phase of EAE. JCI Insight, 7(8), e155563.

+ Open protocol

+ Expand

Gene Expression Analysis Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Gene expression analysis was performed as previously described [11 ] using the following methods: Total RNA
was extracted from cells with TRIzol and reverse transcribed using qScript cDNA
Mix (Quantabio, Beverly, MA, USA) to cDNA, which was analyzed by quantitative
polymerase chain reaction (PCR) with SYBR FAST qPCR Master Mix (Kapa Biosystems,
Wilmington, MA, USA) with primers shown in Supplemental Table 2. Relative mRNA
levels were determined using the -ΔΔCt method
[17 (link)] with internal control,
β-actin gene (Actb). Error bars denote mean ± SEM
of biological replicates in Figures
1C–D, 1F–H, and
6K. In Figures 1A and 2A, B error bars (sometimes too short to be
identified) were calculated using RQ-Min =
2^{−(ΔΔCt +T*SD(ΔCt))} and RQ-Max =
2^{−(ΔΔCt –T*SD(ΔCt))} from triplicate wells as
suggested by Applied Biosystems (manufacturer of qPCR machines). Results shown
are representative of multiple independent experiments with similar results.

Deerhake M.E., Cardakli E.D, & Shinohara M.L. (2022). Dectin-1 signaling in neutrophils upregulates PD-L1 and triggers ROS-mediated suppression of CD4+ T cells. Journal of leukocyte biology, 112(6), 1413-1425.

+ Open protocol

+ Expand

Quantifying Gene Expression in ex vivo Stimulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

To evaluate gene expression in ex vivo stimulation experiments, total RNA was extracted from cells with TRIzol on purified cells, while RNeasy Micro Kit (Qiagen) was used for flow cytometry-sorted cells. cDNA synthesis was performed using qScript cDNA Mix (Quantabio). Real-time, quantitative polymerase chain reaction (RT-qPCR) was performed with SYBR FAST qPCR Master Mix (Kapa Biosystems) with primers shown in Table S1. Relative amounts of qPCR product were determined using the −ΔΔCt method (Livak and Schmittgen, 2001 (link)) comparing relative expression of housekeeping (Actb) and target genes. Error bars denote mean ± SEM of biological replicates in Fig. 4A, B; Fig. 5A; Fig. 7D, F; Fig. S4G–I; and Fig. S7E. In Figures 4D, Fig. 5L, N, O; Fig. S4F; and Fig. S5A–D, F error bars (sometimes too short to be identified) were calculated using RQ-Min = 2^{−(ΔΔCt + T *SD(ΔCt))} and RQ-Max = 2^{−(ΔΔCt − T *SD(ΔCt))} from triplicate wells as suggested by Applied Biosystems (manufacturer of qPCR machines). Results shown are representative of multiple independent experiments with similar results.

Deerhake M.E., Danzaki K., Inoue M., Cardakli E.D., Nonaka T., Aggarwal N., Barclay W.E., Ji R.R, & Shinohara M.L. (2021). Dectin-1 limits autoimmune neuroinflammation and promotes myeloid cell-astrocyte crosstalk via Card9-independent expression of Oncostatin M. Immunity, 54(3), 484-498.e8.

+ Open protocol

+ Expand

Quantitative Gene Expression Analysis of Ex Vivo Stimulated Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

To evaluate gene expression in ex vivo stimulation experiments, total RNA was extracted from cells with TRIzol on purified cells, while RNeasy Micro Kit (Qiagen) was used for FACS-sorted cells. cDNA synthesis was performed using qScript cDNA Mix (Quantabio). Real-time, quantitative polymerase chain reaction (RT-qPCR) was performed with SYBR FAST qPCR Master Mix (Kapa Biosystems) with primers shown in Supplementary Table 2. Relative amounts of qPCR product were determined using the -ΔΔCt method 84 comparing relative expression of housekeeping (Actb) and target genes. Error bars denote mean ± SEM of biological replicates in 5a,d and Supplementary Fig. 6a-d, f error bars (sometimes too short to be identified) were calculated using RQ-Min = 2 -(ΔΔCt + T *SD(ΔCt)) and RQ-Max = 2 -(ΔΔCt -T *SD(ΔCt)) from triplicate wells as suggested by Applied Biosystems (manufacturer of qPCR machines). Results shown are representative of multiple independent experiments with similar results.

Deerhake M.E., Danzaki K., Inoue M., Cardakli E.D., Nonaka T., Aggarwal N., Barclay W.E., Ji R., & Shinohara M.L. (2020). Dectin-1 limits central nervous system autoimmunity through a non-canonical pathway.

+ 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?

Revolutionizing how scientists
search and build protocols!