What is the meaning of zero or undetectable gene expression ct value of RT-PCR?

I'm getting zero or no ct value in my gene expression analysis during RT-PCR analysis. I'm not understanding the meaning of zero ct value, how I should interpret the data now. Should I consider...

02 March 2020 2,976 3 View

Is it ok to use transparent 96 wells plate for h2dcfda assay or I should use black plate with transparent base?

I'm using transparent polystyrene 96 well plate for H2dcfda assay, is it ok or I should use black plate with transparent base.

01 February 2020 5,212 3 View

Which statistical test to perform my MTT dataperformed for different time intervals with different concentrations of a bacterial toxin?

I have treated THP1 and AGS cells for 12, 24 and 48 hours with a bacterial toxin concentrations 0, 5, 10, 20, 40, and 80 ug/ml. Now I want to prove my results with statistical methods but I'm...

10 November 2019 6,437 6 View

Problem during pET15b and pET22b restriction digestion with XhoI and NdeI?

Hello everyone, I'm digesting 2 ug of midiprep isolated pET15b and 22b with BamHI and XhoI at 37 oC for overnight. There is smearing in gel and size of the cut vectors also not appearing actual,...

05 June 2019 1,520 4 View

What would be the conductivity of Tris buffer having molar concentrations 1.5 and 0.5 M with respect to pH 8.5 and 6.5 respectively?

I'm preparing tris buffers of molarity 1.5 and 0.5 M with respect to pH 8.5 and 6.5 respectively. The conductivity of molar concentration 1.5 is 26.0 ms/cm and for 0.5 M is 33.6 ms/cm. Is it a...

03 April 2019 6,071 9 View

How to write short name for downloaded protein sequences quickly to save time?

I have downloaded 1000 protein sequences having very big name in FASTA format. I want to cut short these name in a quick manner. Is it possible through and command line or software or I will have...

03 April 2019 7,274 4 View

Is it good to strip gaps in a protein MSA file for phylogenetic analysis?

I'm analyzing 445 protein sequences of a single protein for phylogenetic analysis. Multiple sequence alignment of these sequences is showing gaps of different sizes. Is it ok to keep these gaps...

03 April 2019 8,456 3 View

How many colonies we should expect after transformation of ligation mixture during a cloning experiment?

I'm getting more than 200 colonies after transforming 2 ul of my ligation mixture from 1;3 ratio. I used 100 ng of vector and now it is going hectic to screen a positive clone because the colies I...

01 February 2019 8,673 4 View

Reason for long streaky colonies after transformation?

Hello every one, I'm getting long streaky colonies after transforming pET15b in DH5alpha. Kindly suggest the reason.

01 February 2019 6,552 7 View

What would be best competent cell to maintain and expression of a secreted Gram negative bacterial toxin gene?

I'm cloning a secreted Gram negative bacterial toxin and not finding any colonies with DH5alpha and pET15b. I'm just wondering what could be the best competent cell for keeping and expression of...

31 December 2018 8,914 8 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

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

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03 March 2021 5,360 2 View

Waters 2707 HPLC autosampler problem?

I am using a 2707 waters HPLC device. When I try to inject a sample, it says missing plate or rack. I changed the needle and calibrated its position but I still get the same problem. I even get...

02 March 2021 1,408 1 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

How to separate water from glycerol solvent in a solvent recovery vacuum distillation column?

I am currently doing my third year design project. I have been assigned to design a glycerol recovery column in vinyl acetate monomer production. The process unit before the recovery column is an...

01 March 2021 9,525 4 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

Any research papers for extraction of nutrient deprivation on macroalgae? Analysed by HPLC-UV and GC/MS. Possibly from macroalgae genus Rhizoclonium?

I am doing my thesis on the nutrient deprivation on the macroalgae Rhizoclonium sp. I will be analysing the levels over a period of roughly 5 days by HPLC-UV and GC/MS. I will be using seawater as...

28 February 2021 6,477 1 View