Why is my GOI not expressing after lentiviral transduction despite RNA increase and protein is detected in transfection?

Hi, I'm trying to create a stable overexpression line with EF1a-FlagGOI-IRES-GFP using lentivirus packaged with pxPAX2 and pM2G. After transduction of 293T with 8ug/ml polybrene, I was able to...

29 January 2021 1,878 4 View

How to correct a deviation in the molar ratio in an Isothermal Titration Calorimetry equipment?

Our equipment of ITC does not reach the molar ratio (N=1) of a titration of EDTA with Ca2+ in MES buffer pH 6. We have cleaned with DCN 90 and HCl pH 2, but the N value is very low, around...

10 November 2020 3,137 3 View

How can I obtain the cohesive energy density and solubility parameter for a crystal using the Materials Studio software?

Hello, I am currently analyzing properties of some minerals such as Bi2O3 by MD using Materials Studio, I need to obtain the cohesive energy density (CED) to obtain the solubility parameter,...

02 November 2020 4,356 1 View

What are the SPSS commands to calculate item discrimination index and difficulty index as part of item analysis?

Hi everyone! I am trying to find the "how to" on item difficulty and discrimination on SPSS and its interpretation. I have read about and performed the command analyze> scale> reliability...

21 September 2020 4,040 3 View

In which solvent should I reconstitute mitoTEMPO?

Dear colleagues, I was wondering which solvent should I use to reconstitute mitoTEMPO, I've seen I could use water or DMSO but I'm not sure about the stability of the compounds in the respective...

18 May 2020 2,882 3 View

Does anyone knows if is there any open source repository of historical aerial photography that includes the north of Chile?

My PhD includes a remote sensing survey of the Atacama Desert using freely available satellite imagery to detect archaeological sites. So far, I have not found any source of historical aerial...

12 May 2020 1,383 8 View

Converting Large Chemical Formats to .pdbqt for AutoDock Vina?

I am trying to convert a large single file library (.sdf, .pdb) containing 50k+ molecules into a .pdbqt file for screening. I have tried using AutoDock Tools but it crashes upon trying to load the...

26 April 2020 5,020 6 View

Do you think the approach to sustainability and environmental issues will change for human society after this pandemic?

We need to be closer to the nature and don't look for money and other values.

29 March 2020 1,238 4 View

Which statistical analysis should I follow for analyzing the effects of two factors in two different cell types cultured in vitro?

Hello all! I have performed experiments involving two different cell types. These two cell types have been cultured in vitro and measurements have been performed after 4, 7 and 10 days of culture....

18 March 2020 6,337 10 View

Why do we use qPCR with a singular reference primer?

Hi, I am doing a qPCR and using a reference primer for my lab research project. However, I am not sure what is the purpose of having a singular primer for my reference gene and having both...

28 February 2020 1,636 3 View

Why Austenite grains in steel show much more twinning than ferrite grains?

It is oftentimes said to be a distinctive metallographic trait that differentiates a fully austenitic and a fully felly ferritic steel without etching that might distinguish between two. Why...

02 March 2021 728 2 View

Samples to study circular dichroism (chirality)?

I want to build a low-cost polarimeter to study circular dichroism (chirality). What type of samples can I use? I am looking for samples that can be found outside the laboratory.

01 March 2021 9,101 3 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

How to calculate the crystallinity of cellulose film?

27 February 2021 2,753 4 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View