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Dithiothreitol (dtt)

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Dithiothreitol (DTT) is a reducing agent commonly used in biochemical applications. It is a small, water-soluble molecule that can reduce disulfide bonds in proteins, helping to maintain their structure and function. DTT is often used in protein purification, electrophoresis, and other laboratory procedures involving the handling of proteins.

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Lab products found in correlation

Ficoll pm 400, by Merck Group (2 mentions) 0.5m edta, by Thermo Fisher Scientific (2 mentions) Sarkosyl, by Teknova (2 mentions) Hw 65s, by Merck Group (2 mentions) Tris ph 7, by Merck Group (2 mentions) Autocad, by Autodesk (2 mentions) Ethylene diamine tetraacetic acid (edta), by Corning (1 mentions)

3 protocols using dithiothreitol (dtt)

1

Colonic Crypt Isolation Protocol

Cited 1 time
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Colonic crypt isolation was performed.48 (link) Briefly, isolated colonic tissues were chelated in buffer consisting of 3 mM EDTA (Corning) and 0.5mM DTT (Teknova) in 1X Dulbecco’s Buffered Saline (DPBS) for 1 hour and 15 minutes rotating at 4°C. The tissue was then transferred to 1X DPBS and shaken rigorously for 2 minutes to separate the colonic epithelium from the tissue. After transfer of crypts to a new tube, shaking was repeated 3X to collect remaining crypts. Crypts were washed in DPBS and divided among various enzymic cocktails and conditions for dissociation.
Arceneaux D., Chen Z., Simmons A.J., Heiser C.N., Southard-Smith A.N., Brenan M.J., Yang Y., Chen B., Xu Y., Choi E., Campbell J.D., Liu Q, & Lau K.S. (2023). A contamination focused approach for optimizing the single-cell RNA-seq experiment. iScience, 26(7), 107242.
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2

Microfluidic Device Design and Fabrication

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Microfluidic devices were designed using AutoCAD (AutoDESK, USA), tested using COMSOL Multiphysics as well as
empirically, and fabricated using soft lithographic techniques22 (link) (Supplementary Data 1). The devices were tested on a Drop-seq setup,
using bare beads (Tosoh, Japan, Cat # DTTiCZIBPBHhf-iF1YcK/" target="_blank">HW-65s) in Drop-Seq Lysis Buffer (DLB7 (link); 10 ml stock consists of 4 ml of nuclease-free H2O, 3 ml 20% Ficoll PM−400
(Sigma, Cat # F5415-50ML), 100 μl 20% Sarkosyl (Teknova, Cat # S3377), 400 μl 0.5M
EDTA (Life Technologies), 2 ml 1M Tris pH 7.5 (Sigma), and 500 μl 1M DTT (Teknova, Cat # D9750), where the DTT
is added fresh) and 1x PBS, to optimize flow and bead occupancy parameters in drops. Droplet generation was assessed under a
microscope in real time using a fast camera (Photron, Model # SA5), and later by sampling the emulsion using a
disposable hemocytometer (Life Technologies, Cat # 22-600-100) to check droplet integrity, size, and bead occupancy.
The device design is provided as a Supplementary File 1 and Supplementary Fig. 1a. The unit in the CAD provided is 1 unit =
1 μm; channel depth on device is 75 μm.
Habib N., Avraham-Davidi I., Basu A., Burks T., Shekhar K., Hofree M., Choudhury S.R., Aguet F., Gelfand E., Ardlie K., Weitz D.A., Rozenblatt-Rosen O., Zhang F, & Regev A. (2017). Massively-parallel single nucleus RNA-seq with DroNc-seq. Nature methods, 14(10), 955-958.
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3

Microfluidic Device Design and Fabrication

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Microfluidic devices were designed using AutoCAD (AutoDESK, USA), tested using COMSOL Multiphysics as well as
empirically, and fabricated using soft lithographic techniques22 (link) (Supplementary Data 1). The devices were tested on a Drop-seq setup,
using bare beads (Tosoh, Japan, Cat # DTTiCZIBPBHhf-iF1YcK/" target="_blank">HW-65s) in Drop-Seq Lysis Buffer (DLB7 (link); 10 ml stock consists of 4 ml of nuclease-free H2O, 3 ml 20% Ficoll PM−400
(Sigma, Cat # F5415-50ML), 100 μl 20% Sarkosyl (Teknova, Cat # S3377), 400 μl 0.5M
EDTA (Life Technologies), 2 ml 1M Tris pH 7.5 (Sigma), and 500 μl 1M DTT (Teknova, Cat # D9750), where the DTT
is added fresh) and 1x PBS, to optimize flow and bead occupancy parameters in drops. Droplet generation was assessed under a
microscope in real time using a fast camera (Photron, Model # SA5), and later by sampling the emulsion using a
disposable hemocytometer (Life Technologies, Cat # 22-600-100) to check droplet integrity, size, and bead occupancy.
The device design is provided as a Supplementary File 1 and Supplementary Fig. 1a. The unit in the CAD provided is 1 unit =
1 μm; channel depth on device is 75 μm.
Habib N., Avraham-Davidi I., Basu A., Burks T., Shekhar K., Hofree M., Choudhury S.R., Aguet F., Gelfand E., Ardlie K., Weitz D.A., Rozenblatt-Rosen O., Zhang F, & Regev A. (2017). Massively-parallel single nucleus RNA-seq with DroNc-seq. Nature methods, 14(10), 955-958.
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