Is my signal sequence is correct or functional?

My cloned gene contain a signal sequence at N terminal . How can I check the nucleotide sequence is a correct signal sequence. The following uppercase letter are signal sequence cloned in...

01 February 2019 7,597 2 View

Can I get single fragment using two PCR product in single reaction ?

I am planing to get two gene together for cloning purpose. My plan is first amplify the gene "x" using a set of primer which will amplify gene x without stop codon. I will amplify another gene "y"...

08 September 2018 6,654 1 View

Can I preserve my Transfected HEK-293 Cells IN -80 degree for few days, Later these will be being used for Microsome Isolation?

Can I preserve my Transfected HEK-293 Cells IN -80 degree for few days, Later these will be being used for Microsome Isolation?

08 September 2018 8,512 1 View

What will be size of my expressed protein If my construct contain Mut. Htt96Q linked with BiRA* ?

My construct contains Htt 96Q followed by BirA* with a 6 amino acid linker between them. Than what will be size of my expressed protein if molecular mass of BirA* is 35.2 KDa? Based on amino...

07 August 2018 3,656 1 View

How long do I need to treat transfected HEK 293 cells with Brefeldin A and the concentration need to see endo H resistance in glycosylated protein ?

I am using knockout HEK 293 cell.

06 July 2018 4,440 2 View

Why I found colony for negative control (contain only double digested vector : one blunt and one sticky end)after transformation in DH5-alfa cells ?

Why I found colony for negative control on selection plate (contain only double digested vector : one blunt (by EcoRV) and one sticky (by SalI) end with ligation mixture, no insert, after...

06 July 2018 231 1 View

I want to ligate my 1kb insert (PCR product) with 6 kb vector, both are digested by ECORV (creating blunt end) and Sal1 (creating sticky end). How?

I want to ligate my 1kb insert (PCR product) with 6 kb vector, both are digested by ECORV (creating blunt end) and Sal1 (creating sticky end). How? Is blunt end may create problem in cloning?

05 June 2018 6,728 3 View

Why Polyst. Flat bot. high Bind. MP (Costar) blank micro-plate is reactive with positive control?

I am using Polyst. Flat bot. high Bind. MP blank (Costar) micro-plate for making ELISA (HIV ), when i am using this blank plate for ELISA assay , it is found reactive with positive control while...

31 December 2016 5,725 6 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

No title

02 March 2021 3,060 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 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

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

No title

28 February 2021 9,936 2 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View