What kind of raw data from NCBI SRA is used for LEfSE?

I don't have any background in bioinformatics just had a basic biostatistic course back in university. If I need to use the package platform like LEfSE to screen for potential biomarkers from...

07 August 2018 3,878 1 View

Any non-OPA HPLC method for amino acids analysis?

I'm planing to analyze amino acids by HPLC. However, OPA-based method is complicated and does not reach the safety policy. Therefore, is there any other HPLC method for amino acids analysis?...

05 June 2018 6,297 10 View

How to minimize a noise background from Sanger sequencing?

I'm working on Sanger sequencing using Bigdye V.3.1 and purify the sequenced product by DyeEx 2.0 kit. The chromatogram was fine and readable, however, it always contains some noise background and...

10 November 2017 9,673 10 View

How to extract the crude enzyme from yeast culture?

I have studied about the mitochondrial enzyme in yeast. So, I want to extract the crude enzyme from cell culture in order to test the enzyme activity. What is the suitable procedure in this case?...

08 September 2016 6,015 3 View

Protein expression in E.coli, located in inclusion-body, how to purify without losing an enzymatic activity?

07 August 2016 766 9 View

I am stuck with a high Tm primer for PCR? Why do I get no PCR product?

31 December 2015 6,655 14 View

What is the suitable plasmid or selective media for S.cerevisiae S288c transformation?

I need to transform pRS416 plasmid containing an interesting point mutation gene to S288c which contained random mutation. Since this strain can grow on SD media so I've just wondered, what media...

01 February 2015 5,198 1 View

How can I verify yeast transformed with point mutation plasmid?

I have introduced 1 base mutation which cause 1 amino acid changed by PCR then transformed this plasmid to DH5-alpha. At this step, I checked desire transformants byspread on selective media...

11 December 2014 8,237 4 View

Buffer to disrupt platelet alpha granules for PF4 quantification?

Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...

03 March 2021 1,499 2 View

Does the pressure(-600 mmHg) affects pH?

When I conducted the degassing experiment using decompression under anaerobic condition(and I used ferrous based chelating agent), the oxidized ferric were observerd. So, could you tell me the...

03 March 2021 4,892 5 View

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 3 View

How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 2 View

Why are my bands weird in this Agarose gel?

Can someone please give me some possible things that could go wrong? Here is my recipe: 0.5g Agarose 50 mL of TAE 1x 1 uL ethyl bromide. Gel was run at 100V for 1 hour. The buffer used is also TAE.

01 March 2021 9,952 3 View

What do I do with DLS peaks that are from the buffer?

I'm looking at the aggregation of my protein sample using DLS. Unfortunately, my buffer (20mM HEPES) also results in a set of peaks. These are at approximately 1 and 1000 d.nm. The lowest peak...

01 March 2021 9,015 2 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

For gelatin coating for culturing HUVEC.Should the gelatin be made in sterile water or PBS? Do I need to autoclave the gelatin soln after making it ?

I want to use 0.1% gelatin. Should that be made in PBS or sterile water? Should I autoclave the gelatin soln after making it in water/PBS?

28 February 2021 274 2 View

The reason that the culture of white fungi Contamination with green mold(Trichoderma), the culture autoclave at121 for 15 min. What is it?

Is the period to autoclave not enough? The inoculation in a hood and flame and UV and alcohol.

27 February 2021 9,356 3 View

Which database would give me this list of chemical components?

Dear colleagues: I would like to know if anyone knows of a database that gives me the parameters pH, K, K, Mg, P, C, N, Zn, Cu, Ni and Cd of common foods. I find a lot of information of proteins,...

26 February 2021 5,525 3 View