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96 well reaction cuvette

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The 96-well reaction cuvette is a laboratory equipment designed for high-throughput applications. It provides a standardized vessel for conducting various chemical and biological experiments in a multi-well format.

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Lab products found in correlation

Tracrrna, by Horizon Discovery (2 mentions) Crrna, by Horizon Discovery (2 mentions) Amaxa 4d nucleofector, by Lonza (2 mentions)

2 protocols using 96 well reaction cuvette

1

CRISPR-Cas9 Editing of Stimulated Treg Cells

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80 μM crRNA (Dharmacon) and 80 μM tracrRNA (Dharmacon) were mixed in a 1:1 ratio and incubated 30 min at 37°C to generate 40 μM crRNA:tracrRNA duplexes. An equal volume of 40 μM S. pyogenes Cas9-NLS (Macrolabs, Berkeley) was slowly added to the crRNA:tracrRNA and incubated for 15 min at 37°C to generate 20 μM Cas9 RNPs. For each editing reaction, 1.5-5x105 stimulated Treg cells were pelleted and re-suspended in 20 μL P3 buffer. 3 μl 20 μM Cas9 RNP and 0.75 μl 100 μM electroporation enhancer (IDT) was added directly to the cells and the entire volume transferred to a 96-well reaction cuvette (Lonza). Treg cells were electroporated using program EH-115 on the Amaxa 4D-Nucleofector (Lonza). 80 μL pre-warmed cRPMI was added to each well after electroporation and the cells were allowed to recover for 30 minutes at 37°C before restimulation.
Schumann K., Raju S.S., Lauber M., Kolb S., Shifrut E., Cortez J.T., Skartsis N., Nguyen V.Q., Woo J.M., Roth T.L., Yu R., Nguyen M.L., Simeonov D.R., Nguyen D.N., Targ S., Gate R.E., Tang Q., Bluestone J.A., Spitzer M.H., Ye C.J, & Marson A. (2020). Functional CRISPR dissection of gene networks controlling human regulatory T cell identity. Nature immunology, 21(11), 1456-1466.
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2

CRISPR-Cas9 Editing of Stimulated Treg Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
80 μM crRNA (Dharmacon) and 80 μM tracrRNA (Dharmacon) were mixed in a 1:1 ratio and incubated 30 min at 37°C to generate 40 μM crRNA:tracrRNA duplexes. An equal volume of 40 μM S. pyogenes Cas9-NLS (Macrolabs, Berkeley) was slowly added to the crRNA:tracrRNA and incubated for 15 min at 37°C to generate 20 μM Cas9 RNPs. For each editing reaction, 1.5-5x105 stimulated Treg cells were pelleted and re-suspended in 20 μL P3 buffer. 3 μl 20 μM Cas9 RNP and 0.75 μl 100 μM electroporation enhancer (IDT) was added directly to the cells and the entire volume transferred to a 96-well reaction cuvette (Lonza). Treg cells were electroporated using program EH-115 on the Amaxa 4D-Nucleofector (Lonza). 80 μL pre-warmed cRPMI was added to each well after electroporation and the cells were allowed to recover for 30 minutes at 37°C before restimulation.
Schumann K., Raju S.S., Lauber M., Kolb S., Shifrut E., Cortez J.T., Skartsis N., Nguyen V.Q., Woo J.M., Roth T.L., Yu R., Nguyen M.L., Simeonov D.R., Nguyen D.N., Targ S., Gate R.E., Tang Q., Bluestone J.A., Spitzer M.H., Ye C.J, & Marson A. (2020). Functional CRISPR dissection of gene networks controlling human regulatory T cell identity. Nature immunology, 21(11), 1456-1466.
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