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Dnase type 1

Manufactured by Thermo Fisher Scientific
Sourced in Germany, Switzerland

DNase type I is a laboratory enzyme that catalyzes the hydrolysis of DNA. It is commonly used in various molecular biology techniques to degrade DNA in order to remove it from samples or reactions.

Automatically generated - may contain errors

2 protocols using dnase type 1

1

Isolation of Immune Cells from Porcine Maternal-Fetal Interface

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The procedure for the isolation of immune cells from the porcine maternal-fetal interface has been described previously (37 (link)). In brief, ME and FP tissues were cut into small pieces and incubated in tissue digestion medium [RPMI-1640 supplemented with 2% (v/v) heat-inactivated fetal calf serum (FCS; Sigma-Aldrich, Schnelldorf, Germany), 25 U/mL DNase type I (Thermo Fischer Scientific), 300 U/mL Collagenase type I (Thermo Fisher Scientific), 100 IU/mL penicillin (PAN-Biotech), and 0.1 mg/mL streptomycin (PAN-Biotech)] for 1 h at 37 °C and constant mixing. Remaining larger pieces of tissue and dead cells were removed by draining the cell suspensions through a coarse-meshed sieve and subsequent filtering through a layer of cotton wool. Suspensions were centrifuged (350 × g, 10 minutes, 4°C), resuspended in 40% Percoll (13 mL, Thermo Fisher Scientific), underlaid with 70% Percoll (13 mL, Thermo Fisher Scientific), and subjected to density gradient centrifugation (920 × g, 30 minutes, room temperature). Isolated leukocytes were washed four times (phosphate-buffered saline (PBS, 2x), RPMI-1640 + 5% FCS (1x), and RPMI-1640 + 10% FCS (1x)) and immediately used for immune phenotyping.
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2

Lung Tissue Dissociation Protocol

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Lung lobes/large chunks from a single mouse were placed into a 2 mL microfuge tube and chopped into small (<2 mm) chunks with sterile scissors. 1 mL of digestion solution (TM Liberase at 4 units/mL (Roche, Basel, Switzerland) and DNAse Type I at 800 units/mL (ThermoFisher) made in sterile PBS) was added to each tube. Tubes were placed on a rotisserie at 37C for 20 min. Digest was then mechanically ground through 100um nylon filters (ThermoFisher) and placed in ACK RBC Lysis buffer (ThermoFisher) for 3 min at RT or as needed. Cells were pelleted and, if further cleanup is needed, a density gradient was used (debris removal solution, Miltenyi, Bergisch Gladbach, GER) according to manufacturer direction.
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