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

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43 views

Hebbian theory “fire together” clarification

Donald Hebb states it as follows: "Let us assume that the persistence or repetition of a reverberatory activity (or "trace") tends to induce lasting cellular changes that add to its stability.… ...
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1answer
68 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 ...
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0answers
122 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 ...
3
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0answers
16 views

Cross section of actin network in neurites

for a simulation I am developing I would like to know how the actin network in neurites is distributed. Is actin confined to the periphery or is the whole neurite shaft containing actin with a rather ...
3
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0answers
55 views

Selection on linked loci in a diploid population

Let’s consider two linked loci $A$ and $B$ that are both bi-allelic. In consequence, we have four different possible haplotypes $A_1B_1$, $A_1B_2$, $A_2B_1$, $A_2B_2$, which frequencies are $X_1$, ...
3
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0answers
34 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 ...
3
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0answers
59 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
62 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
74 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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0answers
54 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
84 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
148 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
39 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
120 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 ...
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0answers
145 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 ...
2
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0answers
82 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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21 views

Has anyone used Crispr/Cas to induce a knock-in in MEF cells?

Does anyone have experience with the crispr/cas9 platform performed on MEF? Or does anyone recall any relevent articles? Thanks
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0answers
16 views

detection of primary origin (natural selection or genetic drift) on alterations in sequences

I recently had a question about how to distinguish the origin of variations in the sequences of rRNA as to whether alterations are generated by genetic drift or natural selection. For me it has not ...
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0answers
25 views

Assembling DNA fragments with ~10kb overlap in vitro

Let's say I have two fragments of bacterial DNA (~50kb in length each) that overlap on the ends by roughly ~10kb; the overlap sequences are unknown. I would like to assemble these into longer ...
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0answers
25 views

Kinesin-5 / cytoplasmic dynein spatial density distribution in neurons

is there some way to experimentally determine the density distribution of those two molecular motors in a Neuron? Fluorescence labeling would be impossible(?) as GFP markers would probably alter the ...
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0answers
29 views

Study case of the inheritance system of Oenothera

I've been told some interesting facts about oenothera. Apparently in this species some lineages have been through some translocations and in results to these translocations and in consequence, some ...
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0answers
98 views

How does a TOPflash/FOPflash assay work to detect beta-catenin protein expression?

I am reading an article (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3596711/) wherein a TOPflash/FOPflash assay is used to detect beta-catenin protein levels in a COS-7 cell line. I can't find a good ...
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0answers
47 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
687 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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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
38 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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564 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
137 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
45 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
49 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
17 views

coevolution by physical interaction between RNA sequences

There are several items that relate to the concerted evolution between pairs of sencuencias (especially structural) rRNA as a consequence of physical interaction in tertiary, or even quaternary ...
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0answers
10 views

border zone of the infarcted heart

I did myocardial infarction experiments on mice and I am a newbie in this area. I need to isolate RNA and protein from the remote and the border zone. Where should I cut the heart so i can separate ...
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0answers
17 views

What models are used for the different pathways of apoptosis?

We are trying to find references for experimental models of different pathways of apoptosis (caspase-dependent and -independent intrinsic apoptosis, death receptor type-1 and -2 mediated apoptosis, ...
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0answers
21 views

Is it possible to synthetically replicate the results of cryptobiosis? If so, how?

Okay if my question seems ridiculous I apologize because I have next to no knowledge of synthetic biology and what the field is capable of doing and it's methods and methodology. Take a organism ...
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0answers
25 views

PQR files make from pdb2pqr are different to GROMACS

I have a pdb file and I would like to make a pqr one. In GROMACS, I use editconf -mead to produce pqr, with topol.top taken from pdb2gmx or use it to produce pdb file, then use pdb2pqr. Which one is ...
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0answers
28 views

What is the suitable terminology to describe this study approach?

I need to know the correct term(s) which are usually used in the parlance of both biology and bioinformatics for this study approach: About 11 transcripts were investigated using qPCR for a number of ...
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0answers
52 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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0answers
15 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
20 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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0answers
19 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
23 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
23 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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0answers
15 views

Can antioxidants abolish Fas- induced apoptosis?

Can antioxidants abolish Fas- induced apoptosis in oral mucosa?
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0answers
57 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 ?