EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Why is my qPCR suddently not working?

I've been working with qPCR for some months now. I amplify 2 genes (PSEN2 and HN1), and the protocols are optimized for both and have been working fine. The other day I amplified HN1 for some...

21 February 2021 3,437 3 View

Can ethanol be used as an analgesic?

If ethanol has analgesic properties, why isn't it used for clinical purposes?

18 February 2021 8,227 4 View

Acid phenol:chloroform (25:24:1) for RNA extraction instead of (125:24:1)?

Hi everyone! I am about to extract RNA for RNA sequencing assays and my Invitrogen kit/protocol...

14 February 2021 1,755 3 View

Are there any public datasets of consumer complaints about service industries at the European level?

06 January 2021 1,718 2 View

No title

16 December 2020 9,657 5 View

Tools for analysis of interaction between lncRNAs and proteins/miRNAs?

Hi! I would like to know what tools I could use to analyze interactions between a new lncRNA and possible protein/miRNA targets. The biggest problem I had to find such tools is that the RNA we...

20 November 2020 8,331 3 View

I would like to ask some questions about concentrations of Urea 3M and 8M in two experiments?

I would like to ask if anyone knows if concentrations of Urea 3M and 8M can interfere with protein purification at Ni-NTA and Glutathione Sepharose 43 resins. Another question, Urea, at 3M and 8M...

17 November 2020 5,014 3 View

I would like to ask if anyone among you knows what is the ideal concentration of TFA to use at a chemical synthesis of peptides?

During a chemical synthesis of peptides by the F-moc method, the following happened: all the initial steps of the synthesis occurred as planned, but in the cleavage, TFA is used. In the lab, there...

02 November 2020 2,997 8 View

Can I use generalized linear mixed model to "fix" data absence?

I have some data of the diet of an omnivorous fish diet collected over 20 years. This data is divided in two seasons, the dry period (drought) and the wet period (flood), and I want to analyze the...

10 October 2020 6,546 6 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Standard curve shifting over time. How do I stabilise the ELISA?

24 February 2021 1,365 3 View

RNA degraded after good nanodrop readings?

23 February 2021 8,850 2 View

How can we determine the positive definiteness of multivariable polyomial? Is there any MATLAB function available for that?

Can we check the positive definiteness of a multivariable polynomial using MATLAB functions? Of course in literature, there are few complex methods that use tensor algebra. However, before going...

16 February 2021 1,218 3 View

How to avoid ATP degradation while performing ATP assay?

I am using the ATP Assay Kit (Colorimetric/Fluorometric) (ab83355) for measuring intracellular ATP from treated cell lines. Although my standards are wokring fine when using the fluorometric...

15 February 2021 3,746 4 View

Dealing with BMDM seeding and RNA extraction ?

biomedical master student here , i am dealing with mice BMDM and would like some help , i seed 0.5 x 10^6 cells/per well in a 6 well plate but when i extract the RNA i get a very low yeild , i...

15 February 2021 1,102 5 View

How long can agarose embedded tissues be kept?

I am embedding PFA fixed tissues in low melting point agarose prior to sectioning and I am wondering how long can these can be kept whilst maintaining tissue integrity and fluorescent signal?...

14 February 2021 6,688 3 View

What is a suitable method to freeze feeders-free, serum free episomal hiPSC?

Hi everyone, I need your professional advice: I started growing episomal iPSC (commercial cell line). I grow them in StemFlex medium. What would you recommend as the most effective freezing...

07 February 2021 8,591 4 View

Freeze/Thaw of LS174t cells?

Hello World, I have been working with LS174t. Complete Growth Media- 10% FBS/EMEM. ATCC says to freeze with 5% DMSO/CGM. when I defrost my vials, they are not attaching to flask. vials put into...

01 February 2021 8,723 2 View