Log In Sign up
The largest database of trusted experimental protocols

Ambion cells to ct kit

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in United States

The Ambion Cells-to-CT kit is a laboratory product designed for the rapid extraction and preparation of RNA from small cell samples. The kit provides a streamlined workflow for isolating RNA directly from cells, without the need for prior RNA purification.

Automatically generated - may contain errors

Lab products found in correlation

96 well plate, by Greiner (2 mentions) Viia7 real time instrument, by Thermo Fisher Scientific (1 mentions) Cfx384 touch real time pcr detection system, by Bio-Rad (1 mentions) Taqman primer probe set, by Thermo Fisher Scientific (1 mentions) Taqman universal pcr master mix, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Cell-to-CT kit (23 citations) Ambion Power SYBR Green Cells-to-CT Kit (4 citations) Ambion Cell-to-Ct kit (2 citations)

5 protocols using ambion cells to ct kit

1

Quantification of Viral RNA Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were seeded on 96-well plates (Greiner Bio-One) and incubated overnight before being infected by designated viruses with the indicated multiplicity of infection (MOI). At designated time points, cell lysates were harvested using the Ambion Cells-to-CT kit (Thermo Fisher) according the manufacturer’s protocol. After reverse transcription (RT) reaction, quantitative PCR (qPCR) was performed. Viral RNA levels were normalized to 18S RNA levels.
The following primers (5’−3’ orientation) were used for the qPCR:
18S-Fwd: AGAAACGGCTACCACATCCA
18S-Rev: CACCAGACTTGCCCTCCA
CV-A10-Fwd: CACTCAACCGGTTTTGGATT
CV-A10-Rev: TAGTTGTGCCACTCGGTCAC
EV-A71-Fwd40 (link): CCCTGAATGCGGCTAATCC
EV-A71-Rev40 (link): ATTGTCACCATAAGCAGCCA
EV-D68-Fwd41 (link): GGTGTGAAGAGTCTATTGAGC
EV-D68-Rev41 (link): CACCCAAAGTAGTCGG
RV-A2-Fwd: ACCTGGCAGATGAGGCTAGA
RV-A2-Rev: TGTTTGGCTTCACACCCATA
RV-B14-Fwd: AGTTGGCCCTGTAACAATGG
RV-B14-Rev: TGCCCAGTTTTTGTTGCATA
RV-C15- Fwd: AATTCTCAGGTGCCATGGAG
RV-C15- Rev: GTTGGGTGATTGCTCATCCT
Note that the downward error bar in Figure 1d, panel d (SETD3KO, 48 hpi) was omitted because it would reach a negative value that is invalid on a logarithmic scale.
Diep J., Ooi Y.S., Wilkinson A.W., Peters C.E., Foy E., Johnson J.R., Zengel J., Ding S., Weng K.F., Laufman O., Jang G., Xu J., Young T., Verschueren E., Kobluk K.J., Elias J.E., Sarnow P., Greenberg H.B., Hüttenhain R., Nagamine C.M., Andino R., Krogan N.J., Gozani O, & Carette J.E. (2019). Enterovirus Pathogenesis Requires the Host Methyltransferase SETD3. Nature microbiology, 4(12), 2523-2537.
+ Open protocol
+ Expand
2

Quantification of Viral RNA Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were seeded on 96-well plates (Greiner Bio-One) and incubated overnight before being infected by designated viruses with the indicated multiplicity of infection (MOI). At designated time points, cell lysates were harvested using the Ambion Cells-to-CT kit (Thermo Fisher) according the manufacturer’s protocol. After reverse transcription (RT) reaction, quantitative PCR (qPCR) was performed. Viral RNA levels were normalized to 18S RNA levels.
The following primers (5’−3’ orientation) were used for the qPCR:
18S-Fwd: AGAAACGGCTACCACATCCA
18S-Rev: CACCAGACTTGCCCTCCA
CV-A10-Fwd: CACTCAACCGGTTTTGGATT
CV-A10-Rev: TAGTTGTGCCACTCGGTCAC
EV-A71-Fwd40 (link): CCCTGAATGCGGCTAATCC
EV-A71-Rev40 (link): ATTGTCACCATAAGCAGCCA
EV-D68-Fwd41 (link): GGTGTGAAGAGTCTATTGAGC
EV-D68-Rev41 (link): CACCCAAAGTAGTCGG
RV-A2-Fwd: ACCTGGCAGATGAGGCTAGA
RV-A2-Rev: TGTTTGGCTTCACACCCATA
RV-B14-Fwd: AGTTGGCCCTGTAACAATGG
RV-B14-Rev: TGCCCAGTTTTTGTTGCATA
RV-C15- Fwd: AATTCTCAGGTGCCATGGAG
RV-C15- Rev: GTTGGGTGATTGCTCATCCT
Note that the downward error bar in Figure 1d, panel d (SETD3KO, 48 hpi) was omitted because it would reach a negative value that is invalid on a logarithmic scale.
Diep J., Ooi Y.S., Wilkinson A.W., Peters C.E., Foy E., Johnson J.R., Zengel J., Ding S., Weng K.F., Laufman O., Jang G., Xu J., Young T., Verschueren E., Kobluk K.J., Elias J.E., Sarnow P., Greenberg H.B., Hüttenhain R., Nagamine C.M., Andino R., Krogan N.J., Gozani O, & Carette J.E. (2019). Enterovirus Pathogenesis Requires the Host Methyltransferase SETD3. Nature microbiology, 4(12), 2523-2537.
+ Open protocol
+ Expand
3

Viral RNA Quantification Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were plated in 96-well plates (in triplicates for each condition), infected with a moi of 0.1 pfu/cell and treated with 8 μM NGI-1 for HEK293FT cells and 4 μM NGI-1 for HAP1 cells. RNA was harvested using the Ambion Cells-to-CT kit (ThermoFisher Scientific) at 48 hours post infection for flaviviruses, 24 hours for CHIKV and VEEV, and 16 hours for poliovirus. After RT reaction quantitative PCR was performed, where viral RNA levels were normalized to 18S RNA levels.
Puschnik A.S., Marceau C.D., Ooi Y.S., Majzoub K., Rinis N., Contessa J.N, & Carette J.E. (2017). A small molecule oligosaccharyltransferase inhibitor with pan-flaviviral activity. Cell reports, 21(11), 3032-3039.
+ Open protocol
+ Expand
4

Quantifying Cardiac Marker Expression in iPSC-CMs

Check if the same lab product or an alternative is used in the 5 most similar protocols
RNA from iPS cell derived cardiomyocytes was isolated using the Ambion Cells-to-Ct Kit (Life Technologies) according to the manufacturer. Cardiomyocytes were dissociated as described above. To quantify the expression of pan-cardiac marker cardiac troponin T (cTnT) and ventricular specific marker myosin light chain 2 (MYL2) during cardiac differentiation, quantitative polymerase chain reaction (QPCR) was performed using pre-designed TaqMan assays (S1 Table). QPCR reagents were assembled in 384-well plates using 10 μL Taqman Gene Expression Mastermix, 5 μL of nuclease free water, 1 μL of the Taqman Gene Expression Assay of choice and 4 μL of cDNA. The PCRs were run on a ViiA7 real time instrument (Life Technologies) using the following program: 2 minutes at 50°C, 10 minutes at 95°C, 40 cycles of 95°C for 15 seconds and 1 minute at 60°C. Relative gene expression was calculated with the ΔΔCt method normalizing target gene expression to B2M [24 (link)].
Fuerstenau-Sharp M., Zimmermann M.E., Stark K., Jentsch N., Klingenstein M., Drzymalski M., Wagner S., Maier L.S., Hehr U., Baessler A., Fischer M, & Hengstenberg C. (2015). Generation of Highly Purified Human Cardiomyocytes from Peripheral Blood Mononuclear Cell-Derived Induced Pluripotent Stem Cells. PLoS ONE, 10(5), e0126596.
+ Open protocol
+ Expand
5

RNA Extraction and qRT-PCR Analysis Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
The Ambion Cells-to-CT Kit (Life Technologies, Frederick, MD, USA) was used to extract RNA and reverse transcribe RNA to complementary DNA for qRT-PCR without having to purify RNA prior to amplification as per manufacturer’s instructions. qRT-PCR experiments were conducted using a CFX384 Touch Real-Time PCR Detection System (Bio-rad, Hercules, CA, USA) and TaqMan Universal PCR Master Mix (Life Technologies, Frederick, MD, USA). The housekeeping genes GAPDH and β-actin were used for ΔΔCt calculations and relative expression was calculated using Taqman primer/probe sets (Life Technologies, Frederick, MD, USA).
Steinway S.N., Zañudo J.G., Michel P.J., Feith D.J., Loughran T.P, & Albert R. (2015). Combinatorial interventions inhibit TGFβ-driven epithelial-to-mesenchymal transition and support hybrid cellular phenotypes. NPJ Systems Biology and Applications, 1, 15014-.
+ 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!