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Questions related from Levi Adams
Note - this is not about Next-gen sequencing or other major deep-sequencing projects I do a lot of simple (cheap) basic sanger sequencing of plasmids with companies like Macrogen and Genewiz....
08 August 2019 6,041 8 View
I'm studying glioblastoma and I can't help but notice that a large majority of the basic research papers use this one cell line. Is there a resource anyone knows about that would allow to to see...
02 February 2018 1,322 3 View
We are trying to do RNA FISH on a human midbrain sample and are having an issue with auto-fluorescence from the neuromelanin naturally present in the sample. Does anyone have a method to remove...
01 January 2018 4,557 0 View
I am studying vasculogenic potential of cancer stem cells, and so I culture stem cell spheres on Matrigel. After several days, they form 3D tube structures into the gel. Inhibition of my protein...
11 November 2017 329 9 View
I have recently run into a few issues with my loxP constructs. Currently I have a floxed DSRed sequence with Stop codons functioning as a 'stutter' cassette for downstream genes. This way, my...
11 November 2017 4,320 2 View
I am beginning to perform several experiments using cancer cells in a hypoxic environment. Our core facility has a hypoxic incubator that uses nitrogen to displace oxygen down to a minimum of 1%...
10 October 2017 236 4 View
I have isolated tumor-initiating (cancer stem) cells from a solid tumor and have hypothesized that our molecule of interest mediates the differentiation of these cancer stem cells into...
08 August 2017 3,709 1 View
I need to very specifically detect hydrogen peroxide (preferably while not picking up superoxide) in cells that express GFP. Because of the GFP, I can't use DCFDA as the spectra overlap. My boss...
08 August 2016 5,589 1 View
I've been trying to subclone a LoxP-DsRed-LoxP insert, and I'm having a lot of difficulty with the PCR. I played with the annealing temp and tried adding DMSO and High GC enhancer, but no...
06 June 2016 4,007 8 View
I'm making a large, complex vector containing an oncogene. I want to limit expression to a specific subset of Cre-expressing cells, so I would like to add a loxP-stop-loxP to my vector. After...
04 April 2016 4,030 7 View
I'm trying to clone several genes into one plasmid, and was looking at using a 2A self cleaving sequence because its short. So: start-Gene A -- 2A-- Gene B - stop As far as the actual cloning -...
11 November 2015 5,045 3 View
I am attempting to design a plasmid that expresses two shRNA constructs under the U6 promoter and a protein under CMV. This limits my vector choice away from many of the commercially multi-shRNA...
10 October 2015 7,231 5 View
