Cell culture is the process of growing and maintaining living cells in vitro.
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What mammalian cell lines are easiest to grow and transfect?
I'm working with HEK293 cells, which are very easy to transfect. Does anyone have other suggestions for cell lines with very high transfection efficiency? Ideally the chromosomes would also be diploid ...
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Copper for cell incubator to prevent contamination
For some reason the lab seems to have a problem with contamination every so often. It's virtually impossible to prevent bacteria, viruses, fungi, etc. from getting into the incubator every time you ...
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Where to put the gene after eukaryotic promoter for best expression levels?
As far as I know there is an optimum distance between a promoter and the gene for the best expression levels. What is that distance for common promoters like CMV, SV40? If you have a first hand ...
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How can I resuspend a cell pellet without harming the bacteria?
When using a preculture with Ampicillin in my protein expression, I have to get rid of the preculture medium to avoid carrying over too much beta-lactamases that will destroy the ampicillin in the ...
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Resuspending Cells from a filter plate
My experiment involves pulling down cells on to a filter plate for a variety of assays. However, I want to ensure that my cells are alive so I am trying to resuspend them so that I can do an accurate ...
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How much time do INS1-E and MIN6 cells need after splitting?
I am currently doing an experiment on cells to test the internalization of a protein.
Normally, I seeded my cells the day before the incubation. This worked well for Hela, CHL or PANC1 cells. However, ...
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Is cell senescence in culture comparable to that in vivo?
A cell is 'senescent' when is has permanently left the cell-cycle. This can be caused by stresses, or by reaching the 'Hayflick limit' (the cell has reached its replicative lifespan, as defined by its ...
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Hoechst or DAPI for nuclear staining?
When is it best to use Hoechst vs. DAPI for nuclear staining? They seem to be very similar on paper. Are there situations where one is clearly preferable?
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High Glucose vs Low Glucose DMEM for Cell Culture
I've noticed that in mammalian cell culture, there are often two types of DMEM available. High Glucose and Low Glucose. Does it matter which type I use for culturing of cells (e.g. Hela or HEK293)? ...
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Tubing connector identification request
this may not be on topic but I'm hoping someone here can tell me the name/manufacturer of the connectors used with the thin teflon tubing in this picture:
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Appropriate methods for triteration?
What is the appropriate method to pipetting up and down mammalians cells to sufficiently break up clumps. This should also keep in mind the issues from Pipetting damage on cells. Visually one can see ...
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What is an efficient way to spike 10-50 cells to a culture
I'm trying to add on the order of 10-50 mammalian cells to a mixture of other cells (order of 1e6 cells). What is the best way to do this?
(edit) I guess this wasn't clear. I would really like to ...
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Why is beta-mercaptoethanol often added to cell culture media
Many protocols suggest that beta-mercaptoethanol is necessary for growing cells. It is a reducing agent but what does it mechanistically do for your cells. When would one not add it.
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Pipetting damage on cells
I'm curious how much damage is potentially inflicted by shear stress by pipetting. I know that with syringes for stem cell injection cause a lot of damage. However, to what extend does this happen ...
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What temperature should mammalian B-Cells be stored at outside of the incubator?
I'm working with murine B-cells. The general protocol is to keep cells on ice to keep them from dying but I've noticed that it makes these cells aggregate and precipitate out. I've heard suggestions ...
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What is the effect of exendin on beta-cells
Do you know if exendin, an analog of GLP-1 (glucagon like peptide-1), can be toxic for beta-cells? For example what is the effect on INS1 or Min6 cells at a certain concentration or after 90 mins of ...
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Providing small molecules to cells on a filter plate
Lets imagine that I have mammalian cells that I've immobilized on a filter. Now I want to keep providing small molecules to these immobilized cells without resolubilizing the cells.
The caveat is ...
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Can a cell culture insert plate be used with a vacuum manifold?
There exists 96 well plate setups for use with vacuum manifolds. Is it possible to use 6/12/24 plate inserts with the same setup?
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How do you break up cell clumps when passaging?
I am working with HepG2 cells, and they really like to form clumps. Pipetting up and down in TrypLE does not seem to be very effective in breaking them up.
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Which human cell lines do not express the GLP-1 receptor?
I need a human cell line that does not express the GLP-1 (glucagon like peptide-1) receptor.
I'm working with HeLa cells, do those express the GLP-1 receptor? Which other cell lines exist that don't ...
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How can I keep HEK cells alive while expressing NMDA receptors?
I am trying to express functional NMDA receptors in HEK293 line cells for single channel recording experiments.
The HEK cells are maintained in the standard way (Thomas & Smart 2005) and ...
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How long does antibiotic-dosed LB maintain good selection?
Various people in our lab will prepare a liter or so of LB, add kanamycin to 25-37 mg/L for selection, and store it at 4 °C for minipreps or other small cultures (where dosing straight LB with a 1000X ...
