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

what is the simplest definition of ion exchange capacity

What this unit identifies for the ion exchange capacity ca. 0.92 meq. g-1. I am a newcomer in the membrane area. I just started. I highly appreciate your help

08 May 2021 0 0 View

what is the simplest definition of ion exchange capacity

What this unit identifies for the ion exchange capacity ca. 0.92 meq. g-1. I am a newcomer in the membrane area. I just started. I highly appreciate your help

08 May 2021 0 0 View

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View

I want to performe MALDI-TOF MS for bacterial identification, I would appreciate it if anyone can tell me the matrix solution preparation protocol ?

based on what I've studied on multiple papers, for peptide and protein identification Alpha-cyano-4-hydroxycinnamic acid (CHCA) is an appropriate matrix to use, I also know that the CHCA powder...

26 February 2021 6,799 3 View

Hi i want to ask that in order to find out LC 50 (96 hours) of particular toxin for fish. Should we keep the animals with food or without food?

if we expose the fish after acclimatization to particular toxin in order to find out LC 50 (96 hours).Should we keep the fish off feed or with feed for that duration. Some literature show that...

24 February 2021 357 2 View

Cloudy/high background western blot - can anyone help me?

Hi everyone! I try to detect VDR protein (~48 kDa) using Western Blot. After SDS-PAGE using 10% MP TGX Stain-Free Gel from BioRad I transferred the protein onto an metOH activated PVDF membrane...

23 February 2021 6,266 3 View

Universal bacterial primers targeting a mRNA?

Hi, I'm looking for universal primers targeting a bacterial mRNA to quatify bacterial load by qPCR in a complexe mixture (eukaryote and prokaryote RNA). Ribosomal RNA being depleted, I absolutly...

22 February 2021 9,969 3 View

Does anyone have any ideas why drug-resistant malaria is increasing in Southeast Asia even when those drugs are no longer used?

When dealing with bacterial toxins, there are three varieties of microbes that develop in your petri dish: toxic, resistant, and susceptible. Initially, there are only toxic and vulnerable...

21 February 2021 6,084 5 View

Protocol for membrane extraction E. coli?

Hello! I am interested in determining whether a poly-unsaturated fatty acid I have engineered E. coli to produce is being incorporated into the membrane of the cells or if it is simply a free...

21 February 2021 7,447 2 View

How to dilute Toxin-DMSO solution?

I have a concentrated solution of a toxin(50 micromolar) in DMSO 100%. I need to dilute it inorder to dilute both the toxin and DMSO wich can be toxic to cells. Which would you guys recommend to...

19 February 2021 7,447 2 View