Hi, Can any one give me equation for measuring the relative and residual activity of an enzyme?

I need to measure the relative and residual activities for my enzyme is there any simple equation to understand how to measure them?

31 December 2016 9,634 5 View

What is the unit of intracellular Reactive Oxygen Species (ROS)?

I measured intracellular ROS for bacterial cells so what will be the unit ? i can say Micro mol/g or what? I'll appreciate your reply.

09 October 2016 5,273 0 View

Why different PCR band appear in gel electrophorysis?

I did cloning for certain insert (1155 bp) into vector (5000 bp) and after transformation i checked the success of transformation by doing PCR for resulted colonies and I got band in the right...

06 July 2016 6,515 9 View

I have DNA template which have 2 restriction enzyme sites how can I change these restriction sites?

05 June 2016 5,013 5 View

Which reference gene primer can be used to normalize qRT-PCR data from E. Coli?

I want to study mRNA expression of my gene at E. coli (I did cloning for the gene from bacillus to E. Coli before), I finished the extraction step for RNA but I don't know which reference gene...

02 March 2016 7,970 4 View

I wanna ask about the difference between relative activity and specific activity?

I also wanna know how to calculate them for laccase i usually calculate laccae activity as U/L.I have attached article for more explanation.

11 December 2015 522 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

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

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

Can random sampling have inclusion criteria?

As random sampling means that everyone in the population have an equal chance to be chosen, can it have inclusion criteria? My research participants have been chosen based on the inclusion...

26 February 2021 6,905 10 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

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View