The study of the molecular processes of the nucleus and cell function.

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98 views

Image analysis of GFP-tagged protein localization bursts

I'm reading an article (full text here) that analyze the dynamics of localization of a GFP-tagged transcription factor (Crz1) over the time at the single-cell level, by taking movies in a fluorescent ...
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2answers
48 views

What is the impact of prolonged electromagnetic exposure on human health?

What are the health effects of being exposed to a very strong electromagnetic field caused by two resonating superconductive coils (not static magnets) for a period of years?
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101 views

What is the fastest way to build an alanine scanning library?

For interfacial studies, I would like to build an alanine scanning library for one of my proteins examining 20 sites. I will ultimately express the gene using E.coli cell-free protein synthesis. I ...
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0answers
54 views

What are the costs associated with carrying lots of genetic material

What are the costs (if any) associated with carrying lots of genetic material? energy for copying? raw material for copying? space in the cell? Maintenance cost (matter and energy)? time to copy is ...
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0answers
37 views

What are the lifetimes of cellular components on organelle or molecular level?

I was thinking that even though I know how generally the cell works, I don't really have a feeling of how volatile it is. I want to know what is the lifetime/turnover rate of different constituents of ...
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0answers
30 views

Adenosine metabolism

Are adenosine or its catabolites increased in inflamed airways? How can I assess this? I am trying to use inhibitors for adenosine deaminase, xanthine oxidase, and purine nucleoside phosphorylase, but ...
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0answers
48 views

Large scale reverse transcription?

I need to make RNA:DNA duplexes. I can make 100 to 200 ug of mRNA through in vitro transcription, and I know how to use reverse transcription to make a cDNA library, but I have questions with this. ...
3
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0answers
54 views

Will eukaryotic RNA fold in the same way in prokaryotes?

As far as I know, there are no specific eukaryotic or prokaryotic factors that aid in RNA folding other than cellular environment (salt and ion concentrations, dissolved molecules, etc). Are there any ...
3
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0answers
68 views

How does LCR compare to Assembly PCR

The question pretty much explains itself. How do the two methods compare? I've always used Assembly PCR but the method is prone to mistakes and I'm curious how it compares to Ligase Chain Reaction ...
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21 views

Gibson assembly using NeBuilder

I am supposed to construct a plasmid that contains features from two other plasmids. My strategy is to generate three fragments form the two other plasmids. I was encouraged to try Gibson assembly, ...
2
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0answers
39 views

RNA isolation from Drosophila head

I need to isolate RNA from Drosophila head. I basically chop the head off and first homogenize it with a homogenizer (similar to this: ...
2
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0answers
54 views

The mechanism of mechanoreception?

I am interested in knowing the molecular mechanism behind mechanoreception/mechanotransduction (i.e. mechanism behind receptor potential generation on mechanical stimulation). I know that most ...
2
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0answers
110 views

Image Processing Suite for bacterial microscopy: Schnitzcells or MicrobeTracker?

I am looking to start doing some work tracking the size and growth of individual bacterial cells in the microscope. In order to analyze the images I need software that can segment the cells, ...
2
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0answers
35 views

Shortest route for a Nitrogen atom from fossil fuel to a protein in your body?

Millions of years ago fossil fuels originated from plants. A nitrogen atom from fossil fuel is now inside in a protein molecule in your body. What is the shortest path that the nitrogen atom ...
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0answers
87 views

Can Pfx polymerase add only one 3' A overhang?

I am trying to clone a PCR product that was amplified using Pfx polymerase into pGemT vector. I had to A-tail the PCR product using Taq polymerase since Pfx only generates blunt end products. My ...
2
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0answers
109 views

Can I heat Trizol?

I wonder if I can heat Trizol reagent for 30 min 65C. The goal is to disrupt protein-RNA complex while inhibiting nucleases. (I can't use RNasin cause it's inactivated in 65C, and can't use RVC cause ...
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0answers
65 views

Optimization of annealing temperature

I am trying to optimize annealing temperature for some primer pairs. I have tried optimization using cDNA, genomic DNA, Taq polymerase, phusion polymerase etc., but every time I am either getting ...
2
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0answers
80 views

Mouse meta-globin mRNA problem

This is an mRNA strand of mouse meta-globin: 5'-ccccagauacggaauucgaau-3' A) Which small RNA (below) is most likely to regulate expression of meta-globin? ...
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0answers
41 views

DNA content in seeds vs. fruit flesh

Is there a publication comparing DNA yield and / or PCR-amplifiability after extraction from fruit flesh (like apples, oranges, cherries etc) in comparison to seeds of the same fruits ? I would prefer ...
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0answers
465 views

Bacterial 16S rRNA PCR amplification with universal primers

I am doing an experiment to amplify the 16S rRNA gene from bacteria present in gut contents of fish. After extracting DNA, I perform one-step PCR with universal bacterial primers (27F, 1492R) and I ...
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0answers
35 views

Can DNA ploidy analysis differentiate field cancerization effects of oral potentially malignant disorders?

Can DNA ploidy analysis differentiate field cancerization effects of oral potentially malignant disorders [ Leukoplakia, erythroplakia, oral submucous fibrosis, etc] ?
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0answers
37 views

Protocol for immobilize oligo to solid

I'm learning about microarrays and understand that an oligonucleotide can be immobilized to a solid like a CD acrylic plastic by modifying the oligo. Does anyone have a good protocol for modifying a ...
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0answers
34 views

Are stable proteins tagged by ubiquitin?

What would happen if an extremely stable protein was created because of some error? Would that protein be tagged by ubiquitin?
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0answers
440 views

Low RNA yield, low 260/230 ratio

I extracted total RNA from animal tissue using the Qiagen RNeasy kit, however my RNA yield was extremely low and the 260/230 ratio was around 0.3. This is the protocol I followed: The animal ...
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0answers
104 views

What mammalian cell lines are easiest to grow and transfect?

I'm working with HEK293 cells, which are very easy to transfect. Does anyone have other suggestions for cell lines with very high transfection efficiency? Ideally the chromosomes would also be diploid ...
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0answers
44 views

Experience with Detox Cloning

I just saw this paper. A Highly Efficient Molecular Cloning Platform that Utilises a Small Bacterial Toxin Gene I was curious if anyone had experience with this cloning strategy using the IbsC ...
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0answers
47 views

Solid phase use in HIV rapid tests

I have another question in regards to my HIV test research. The rapid tests like Orasures Oraquick contains a strip of synthetic peptides that are used to represent proteins found in the envelope ...
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0answers
44 views

Are DNA molecular motors described by cellular automata rules?

While watching this Molecular Visualizations of DNA it struck me how much the motion resembles Conway's Game of Life. And not only visually; each molecule/atom is described by a finite set of ...
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14 views

Testing gene against resistance and susceptible cultivars

I have one gene cloned from plant fungal pathogen using pure culture. Now I want to characterized this gene using molecular methods for which I have few resistance and susceptible cultivars of the ...
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0answers
16 views

How do you express and detect GLP-1 receptors?

I am currently working with a peptide which is an analogue for glp-1, but during invitro studies am not able to detect for the presence of GLP1- receptors. The cell line used is Min-6. How do I detect ...
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16 views

Does biotin affect downstream experiments, such as RT-qPCR?

I have two biotins--one of which is binding my RNA with cleavable bonds, and the other with irreversible bonds. Does biotin have an effect if it keeps its bonds with RNA? If it does, why is this the ...
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0answers
20 views

How do you amplify a specific AFLP marker?

I'm trying to clone a specific AFLP marker out of Medicago truncatula. I know the selective bases I need for the end of the primers. My problem is that all the AFLP cloning kits I have found are ...
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0answers
25 views

Do you know a learning by research platform for biology?

I work in a university in Europe and I'm looking for new communities of biology enthusiasts. I'm very interested in the "synthetic biology" field and I'm searching good, innovative, if possible ...
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0answers
27 views

What is a phospho-protein binding domain?

Is this just a domain that binds proteins that have been phosphorylated? And it mediates signalling between an activated/phosphorylated protein? How is this significant with BRCA1?
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19 views

Where can I get yeast mutants?

I want to use some yeast mutants to conduct gene functional analysis, and the gene was isolated from plants. Can anybody give me some information regarding the websites or agencies from which I can ...
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15 views

Can antioxidants abolish Fas- induced apoptosis?

Can antioxidants abolish Fas- induced apoptosis in oral mucosa?
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43 views

Realtime PCR plate cleaning

Have anyone used realtime PCR, is that the plate for realtime PCR is one-time using or not ? Otherwise how to clean the plate after reaction to prevent wrong result in next time using ?