Gowthami Mahendran

6 Questions 12 Answers 0 Followers

Questions related from Gowthami Mahendran

Is it necessary to change media after transfecting cells with Lipofectamine3000?

I transfected HCt116 cells with GFP using Lipo3000. I didn't change media after transfection as the manufacturer guidelines say not necessary. However, I noticed a change in the cell morphology...

29 April 2023 6,047 6 View

Is loading equal volume of protein in SDS PAGE correct?

I am comparing the protein expression levels in subcellular fractionations (Cytoplasm and nuclear). If I load an equal volume of both extracts (15 ul each to individual wells), is this still valid...

27 January 2023 6,229 3 View

What is the right media for HCT116?

Is DMEM or DMEM/F-12K the right media for culturing HCT116 cells apart from McCoys media? Did anyone use the gibco one? I see a lot of dead cells when using DMEM. It would be helpful if someone...

21 June 2022 957 3 View

How is FPKM calculated from raw gene count file?

I am working with RNAseq and qRT-PCR validation results to see a correlation. I am comparing a diseased group to a healthy group of cells. I have 4 biological replicates in each category. However,...

04 June 2022 7,964 5 View

BN-PAGE gradient gels?

I have tried using BN-PAGE for mitochondrial complexes isolation. I have followed several protocols (Nature protocols-2006, JOVE-2017). But I experienced a problem with running the gel. The dye...

27 June 2020 5,587 6 View

What is the purpose of adding detergent during sample prep for the BN PAGE and how to find the volume of it for mitochondrial complex separation?

I am working with BN PAGE for the first time and I am not sure about the amounts of detergent (digitonin) that should be added. Also, I am looking for methods to find the concentration of the...

17 March 2020 2,240 9 View