Rosa Aledo

9 Questions 23 Answers 0 Followers

Questions related from Rosa Aledo

What it is the best method to transfect mirna mimics and UTR plasmids to make a luciferase assay?

I would like to know a goog transfection reagents to use with plasmids and mirna mimics together, looking the different lipofectamines I wonder if it is necessary to use for example the lipo 3000...

05 May 2017 7,171 2 View

What it is better to amplify 3'UTR from genomic sequences of by RACE using RNAs?

I would like to clone my whole 3'UTR in a luciferase vector and I was wondering what it is better to use genomic sequences to obtain by PCR or to use RNA to obtain by RACE i I was reading that...

12 December 2016 314 0 View

What is the best and easy software to analyze promoters?

I used genomatyx to analyze transcription factors in promoters sequences, but now I have many problems to login, so does anyone knows another facility to do this?

10 October 2016 6,030 5 View

Too much beta lactamase in protein expression BL21 cultures?

Trying to express recombinant GST-proteins after overnight culture at 18ºC with 0.1 mM IPTG I observed also with my fusion protein a band corresponding to beta-lactamase after maldi analysis. How...

07 July 2016 9,479 5 View

Why is recombinant mutant protein expressed less than wild-type?

I express a recombinant proteins in E. coli (wild type and mutant), the mutant one is much less expressed than the wild type and the culture is slower (time to attain DO 600 of 0.6 before to add...

10 October 2015 1,260 7 View

Bands in EMSA gel remain as a big spot, not well resolved, is it impossible to see single bands?

Could anyone bring me a solution?

10 October 2013 1,255 0 View

Bad resolved band in EMSA?

Sometimes it is not possible to see single bands in EMSA, and complexes remain as a big spot not resolved. Could anyone explain to me how to find a solution?

10 October 2013 3,630 5 View

Protein is not visible on western blot of transfected cells by confocal microscopy

I made cDNA constructions to produce proteins with GFP or dsRED (clontech), and one construction with dsRED after transfection in HeLa cells. I obtained RNA by qPCR, red cells by confocal...

01 January 2013 963 14 View

How can I increase release of my GST-fused protein after digestion with Prescission?

I bind my fused GST-protein to glutathion beads and I cut with Prescission at pH 7.5 to eluate overnight in batch method, sometimes my protein (always the same clone) remains attached to beads...

01 January 1970 6,416 5 View