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

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27 views

When did CRISPR/Cas9 evolve and what is the likelihood that a superior system for live cell genome editing has already evolved on earth since then?

I've read that CRISPR/Cas9 is currently being implemented and tested for its ability to edit genomes in live cells (Summer '14), and that it is supplanting other genome editing tools in the lab, such ...
2
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1answer
34 views

What is a protective epitope?

What is a protective epitope? An epitope is basically a part of antigen. So does it mean that when the epitope combines with an antibody, it helps in the functioning of the antibody instead of going ...
2
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0answers
31 views

What does the term 'epitope mapping' mean? [on hold]

Epitope mapping means identifying the binding site of antibodies on the target antigen. This means that the site to be identified is part of the antigen and not antibody, am I right?
7
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1answer
95 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 ...
6
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2answers
82 views

Why does replication require primers while transcription does not?

In transcription, there is no need for any primer. I guess the basic mechanism of DNA polymerase & RNA polymerase is the same. So why does replication have the need for a primer?
2
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2answers
95 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 ?
2
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1answer
38 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 ...
2
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1answer
51 views

Using RNA-seq to compare gene expression across patients instead of between Control and Experimental conditions

I am working with RNA-seq data from the Cancer Genome Atlas TCGA and I have been reading about how people have compared gene expression levels measured by RNA-seq. Many of the papers I have read talk ...
3
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1answer
38 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 ...
6
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1answer
37 views

What regulates the timing of the motion of molecular machines during DNA Replication?

This question is about this video I found on Youtube. I just want to know what is the mechanism which regulates the timing of motion of the parts of these molecular machines. I know that those big ...
2
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1answer
30 views

What is the biding site code recognized by the parts of the spliceosome

Another question about another Youtube video. At 0:50, the splicing process begins to remove the non-coding section of the DNA (intron), so the different parts of the spliceosome attach to the borders ...
3
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0answers
12k views

Does CPH4 exists? [closed]

I just see the film Lucy, it talks about a molecule called CPH4 which increase human capabilities. Of course it's a film and so it's not scientific facts. During an interview the producer says that ...
2
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1answer
39 views

Why do oncogenes show genetic dominance?

As we know that tumor suppressor gene causes cancer only when both the alleles are recessive in nature.But in case of oncogenes if only one allele is dominant it can cause cancer.Why in case of ...
3
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2answers
454 views

On which strand does the promoter sit?

My book keeps giving different indicators as to whether the promoters are on the coding or template strand. It says the -35 region in prokaryotes must be on the coding strand. It also mentions, that ...
2
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4answers
77 views

determine longest poly-purine tract

how to determine/ find the longest poly-purine tract in any genome and this needs to be on the E. coli genome . is it to figure out the polypurine tract and then figure out the longest chain ? or is ...
3
votes
1answer
53 views

Consumption of NAD+ in glycolysis

Out of 10 steps in glycolysis, only one reaction- Glyceraldehyde 3-phosphate (G3P) to 1,3-bisphosphoglycerate (PGP), uses NAD+ and thereby producing NADH. Furthermore, this very step is solely ...
7
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1answer
79 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 $X_1$, ...
16
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3answers
1k views

How and where, in the human brain, are memories stored?

Background I am a computer programmer who is fascinated by artificial intelligence and artificial neural networks, and I am becoming more curious about how biological neural networks work. Context ...
3
votes
2answers
263 views

How are DNA segments selected in PCR?

I understand that in PCR we're able to amplify only selected portions of the DNA... however despite reading it from multiple sources, I cannot figure out how this selection actually takes place. I ...
2
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1answer
31 views

Why does the pET- expression vector contain a LacI gene additionally to the one in the genome?

The pET plasmid is used for protein expression with T7 promotor in expression strains, such as E.coli BL21(DE3) It contains a lacI gene which codes for the lac repressor protein, a protein of ...
3
votes
1answer
171 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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4answers
83 views

Theoretically, what technique would one use to modify a virus so that it only affected a subset of the population?

I'm writing a novel and i would like to know some of the equipment and techniques involved with modifying a virus. Is it feasible for a virus to be engineered to only affect certain people? It ...
5
votes
1answer
60 views

No digested product bands, but markers are visible. What could be the reasons?

I had to digest my insert containing plasmid and another vector into which my insert has to be ligated, each with BamHI and NotI. Upon digestion, I ran the gel to check for results. The digestion ...
4
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1answer
82 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 ...
5
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3answers
63 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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0answers
25 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 ...
4
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2answers
40 views

What are some atypical examples of positive transcriptional cooperativity?

Cooperativity in gene expression is an important feature of many regulatory networks. Described using the Hill function, the most common example is a transcription factor (TF) that when bound to its ...
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0answers
32 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
5
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1answer
120 views

High frequency human genetic oscillators?

The most well studied genetic oscillators in human genomes are involved in regulating the circadian clock (which operates on an approximately 24-hour cycle) and cell cycle activity (with single cycles ...
1
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1answer
62 views

RNeasy Mini Kit low 260/230 ratio — can I purify this RNA for further use?

I used Qiagen's RNeasy Mini Kit to isolate RNA from 5*10^5 C28/I2 (immortalized human chondrocytes). However, my yield is low (~25 ng/ul), but my 260/280 ratio is great (~2.3), and my 260/230 ratio is ...
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0answers
19 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
17 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 ...
2
votes
1answer
542 views

Lyonization and X-linked disorders?

Lyonization or X-chromosome inactivation is the conversion of all but one, X-chromosomes in Females into non-coding heterochromatin (i.e. deactivated) leading to the formation of one or more Barr ...
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0answers
157 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 ...
3
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1answer
109 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 ...
3
votes
1answer
29 views

Efficiency of plasmid DNA isolation from frozen E. coli cell cultures

Has anyone isolated plasmid DNA from frozen (at -20degrees) E. coli cell cultures (not pellets)? Has that worked and if so, with what yields? What would be the quality of the isolated plasmid DNA if ...
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0answers
28 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 ...
0
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0answers
11 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 ...
1
vote
3answers
42 views

FRET only works for interactions between 1nm to 20nm.

FRET only works for interactions between 1nm to 20nm. How can you be sure that the interaction that you want to study isn't less than 1 nm/greater than 20nm apart?
2
votes
1answer
25 views

Heterochromatin production limitations

Currently playing with some ideas for a project and needed some guidance. I am wondering, both in Drosophila melanogaster and in general, is the amount of heterochromatin a cell/nucleus can produce ...
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0answers
26 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 ...
2
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3answers
97 views

What equipment would one use to modify a virus? [closed]

Would anyone be so kind as to give me a few examples of the equipment involved when modifying a virus? Otherwise my novel may end up reading Eva entered the lab and modified a virus. Not much back ...
6
votes
1answer
242 views

How tolerant are ants to cold?

There was a trail of what are commonly known as sugar ants (small, brown, hyperactive) in my kitchen. Three of them walked onto an ice tray placed in their path. They only walked a short distance on ...
4
votes
1answer
74 views

Do mitochondria use ATP in order make another ATP from glucose?

Do mitochondria use ATP in order make another ATP from glucose, or does it use another source of energy to convert glucose to ATP?
0
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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, ...
6
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1answer
867 views

Why is ATP the preferred choice for energy carriers?

Why is ATP the most prevalent form of chemical energy storage and utilization in most cells?
0
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0answers
29 views

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

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 ...
6
votes
1answer
85 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 ...
7
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1answer
1k views

Difference in length of Okazaki fragments

The length of Okazaki fragments in the lagging strand is about 100-200 nucleotides in eukaryotes and about 1000-2000 nucleotides in prokaryotes. What (molecular mechanism, enzyme type ) ...