Is my signal sequence is correct or functional?

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Could any one please provide me a mammalian expression vector with AviTag to add at C and N terminal of protein ?

I want to add AviTag with my protein at N and C terminal and to express it in mammalian cell line.

09 October 2018 6,660 0 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?

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

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

How to increase a model accuracy ?

dear community, my model is based feature extraction from non stationary signals using discrete Wavelet Transform and then using statistical features then machine learning classifiers in order to...

03 March 2021 6,994 5 View

Which SAR dataset to use for generating coherency matrix and performing target decomposition (Description) ?

Working on chandrayaan-2 DFSAR data, there are three datasets available: 1) Slant range image data product: The slant range complex image file. Each pixel is represented by two 4-byte floating...

02 March 2021 8,481 3 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 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