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I was trying to Immunoprecipitation Drosha protein from mice C57 BJ/6 liver. I lysis 80mg in 1.2ml lysis buffer(from thermo +proteinase inhibitor cocktail). However, after Immunoprecipitation by...

05 June 2018 7,613 3 View

What's the possible explanation for inconsistance between luciferase activity and in vivo overexpression?

09 October 2017 3,883 3 View

Does anyone know any database we could use to search profile of protein expression level of the C2C12 cell line?

I want to know the protein expression profile of C2C12 cellline, which may help me to analyze some problem occured during experiment.

11 December 2014 4,928 3 View

Has anybody else encountered problems with BAC extraction during 3C protocol?

Hey guys, I was doing BAC Clone extraction for Chromosome Conformation Capture(3C). We used alkaline lysis method to lysis the bacterium. Then centrifuged and applied supernatant to DNA extraction...

11 December 2014 2,294 6 View

How can I improve the yield of BAC?

Dear friends, I have a puzzle during BAC extraction. The sample we extracted seems to be contaminated by RNA, although we digested it with RNase A and we cannot see the band for BAC in...

11 December 2014 5,574 2 View

What is the possible explanation for the different outcome between Electrophoresis and Nanodrop 2000?

09 October 2014 9,637 12 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Which fluorescent dye to use for measuring total ROS production?

Is DCFDA appropriate for measuring total ROS production, or can it only detect certain types of ROS? If I want to measure total ROS production, which dye should I use?

01 March 2021 9,447 3 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

How vaccines of covid 19 work?

Millions ask how vaccines of covid 19 be successful and safe, its here to explain generally how vaccine of pfizer work ,mRNA vaccines teach our cells how to make a protein that will trigger an...

28 February 2021 2,723 4 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

The reason that the culture of white fungi Contamination with green mold(Trichoderma), the culture autoclave at121 for 15 min. What is it?

Is the period to autoclave not enough? The inoculation in a hood and flame and UV and alcohol.

27 February 2021 9,356 3 View