Inconsistent results after plasmid re-purification from frozen stock?

Hello everyone, I am performing reporter gene assays on some uORF mutants vs my WT 5' UTR of my gene of interest. I am doing double transfection on HEK293T cells, using Nanoluc as a test reporter...

22 November 2020 2,746 5 View

Forest plots of meta-analysis with odds ratio?

I have found 5 included studies for my meta-analysis with binary outcome. Two of them present the results as n/N (n: events; N: non_events patients), while the other 3 three as OR (95% CI). Is...

09 April 2020 9,315 7 View

Which wearable (e.g. smartwatch) should I use to get real-time fatigue/sleepiness scores?

I want to measure fatigue/sleepiness while driving with a wearable that provides a fatigue/sleepiness score to the drivers in real-time. Therefore, I cannot use devices that collect data that...

15 March 2020 3,527 4 View

Can I simulate IoT device activity using Honeypots or any other Tool?

Hello, I need to compare two different states of performance for IoT devices. State 1: Under Attack State 2: No any kind of attack. I can easily use honeypots to simulate the incoming traffic...

06 March 2020 6,947 3 View

DGGE bands enhancement?

My project concerns analysis of the microbial community of sediments from aquaculture systems in concrete tanks supplied with fresh sea water. We followed a molecular approach through PCR cloning...

25 July 2018 943 2 View

Anyone know how to do GC peak separation?

10 December 2014 7,644 26 View

How has technology impacted the factors of the instructional process, e.g. in the 'Berlin Model' & its phases, e.g. in the 'ADDIE Model'?

23 August 2014 1,029 4 View

What is your experience/opinion/reference regarding code-switching in education?

01 August 2014 4,809 59 View

Rehydration buffer for 2D samples.

I am using the following rehydration buffer for my 2D samples. Do you think any of the components may be altered/ become unstable after long period of storage at -20oC and thus affect the...

05 December 2012 9,738 6 View

Is it possible to freeze an antibody mix?

Is it possible to freeze a primary and secondary (Jackson ImmunoResearch) antibody mix several times and reuse it? Will the antibodies still retain their same affinity after a long period of time?...

15 October 2012 4,955 17 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 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

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

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

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View