Traditional DNA/RNA isolation kits can be used to prepare genomic material for sequencing (RNA-seq, scRNA-seq, WGS, CHIP-seq etc). For some applications with large organoids, tissue specific DNA/RNA extraction kits are recommended. TRI Reagent (T9424) can be added to the dissociated organoids before RNA extraction and can be stored at -80 °C until future use.
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About This Item
form
liquid
technique(s)
cell culture | stem cell: suitable
General description
• Culture of primary, stem cells and cell lines in applications that require reduced or serum-free medium.
• Optimization of transfections that require reduced or serum-free conditions.
• Production of conditioned medium in reduced serum or serum-free conditions.
• Optimized media for the culture and expansion of multiple 3D organoids and tissues systems.
DMEM/F12 PLUS is a proprietary formulation that contains human serum albumin (HSA), defined lipids, trace elements, chemicals and proteins essential for optimal cell growth in reduced serum conditions. The media contains high-glucose and does not contain L-glutamine or penicillin-streptomycin.
Application
Stem Cell Research
Preparation Note
Analysis Note
Osmolality: 290-330 mOsm
pH: 7.2-7.4
Sterility Tested: No Growth/Pass
Endotoxin: <2 EU/mL
Mycoplasma: Negative
Disclaimer
1 of 1
This Item | |||
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| form liquid | form liquid | form liquid | form liquid |
| technique(s) cell culture | stem cell: suitable | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable |
Storage Class
12 - Non Combustible Liquids
wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
Certificates of Analysis (COA)
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How Do you Extract RNA/DNA from Organoids in Cultured in Matrigel?
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How are organoids cultured?
1 answer-
Once isolated, organoids are cultured in specialized media embedded in a rich extracellular matrix (ECM) basement membrane extract hydrogel such as Growth Factor Reduced Matrigel. Media is replenished every other day and cells are passaged once per week (7-12 days) before cells become too large or necrotic. Cells can be passaged in either small clump fragments or as single cells using mechanical dissociation/enzyme-free passaging reagents. Depending on organoid type and confluency, split ratios of roughly 1:3-1:4 can be used when passaging organoids. For sensitive organoid types, ROCKi (Y27632) (SCM075) can be added during passaging to help in promoting cell viability.
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How do you stain organoids with antibodies?
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Organoids can be stained with antibodies for immunocytochemical (ICC)/Immunohistochemical (IHC) analysis using either whole-mount or paraffin embedding/sectioning techniques. It is critical to use antibodies prequalified for organoid analysis.
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