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

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Does mung bean nuclease cleave a phosphate group when it's chewing off 5' or 3' ssDNA ends?

I'm looking to create blunt ends from sticky ends with mung bean nuclease for subsequent ligation. Does anyone know full mechanism by which mung bean nuclease will do this? In particular after the ...
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23 views

What are host cellular factors?

With respect to this paper: Global Analysis of Host-Pathogen Interactions that Regulate Early-Stage HIV-1 Replication What does the term "host cellular factors" mean??
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648 views

What are some of the general characteristics of the DH5 alpha strain?

I can not find some useful sources unfortunately. Please tell me about some important characteristics of DH5 alpha. What makes DH5 alpha suitable for the gene cloning?
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26 views

epitope prediction/ mapping

B-CELL EPITOPE PREDICTION Regarding this article: "Such a molecule can be synthesized or, in case of a protein, its gene can be cloned into an expression vector."----- is a particular line in ...
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44 views

Immunodominant Peptide

Can someone please explain me the concept of Immunodominant peptide in simple language?? I did read the wiki article but did not understand it clearly. Please help! (I have not studied biology since ...
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290 views

Single stranded DNA in gel electrophoresis

From http://www.nfstc.org/pdi/Subject05/pdi_s05_m01_01.htm : ... The separation medium contains a denaturant in order that the electrophoresis is conducted on single-stranded DNA fragments. ...
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59 views

H O N C alternatives for organic life?

Most organic structures built from hydrogen, oxygen, nitrogen and carbon. They are represent 1-,2-,3-,4- bond relations (chemical valence) which allow to build variety of chain structures. Are there ...
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83 views

What are the Gateway clonase enzymes?

The Gateway cloning system utilizes what Life Technologies refers to as "Clonase enzyme mix" to catalyze the BP and LR reactions. What is in this enzyme mix? Is there a sequence for them?
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64 views

Do you have experience with PacBio?

I prepare a experiment and I found $PacBio SMRT$ as the great way to sequence my PCR products. I find the cost: library preparation 655 dollars + sequencing 435 dollars. It seems very low. Do you have ...
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63 views

Would expression of GroEL and GroES in erythrocytes be a potentially effective therapy for sickle cell disease?

Would expression of GroEL and GroES in erythrocytes be a potentially effective therapeutic strategy for sickle cell disease? Why or why not?
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2k views

Explanation of the terms “downstream signaling” and “upstream signaling”

In molecular biology, what's the meaning of the terms "downstream signaling" and "upstream signaling"? What's the difference between them?
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107 views

DNA ligase mechanism

I am unable to understand what happens to the phosphates from ATP. What are they used for ?Charging 3' end or 5'end ?
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244 views

Why did Fair Meiosis evolve?

How and why did Fair Meiosis evolve? I can hardly think that it provided a fitness advantage to the individual carrying the mutation. Why would it? Or did it evolve through lineage selection? Or was ...
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61 views

How do cells figure out the big-picture shape?

I'm wondering what makes cells to divide (and stop) in such a way that they make our hands the shape that our hands are...
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1k views

Why is Sanger sequencing inferior for detecting SNPs in cancer cells?

I am familiar with Sanger sequencing, but at the level of an undergraduate. A lecturer of mine tried to describe Sanger sequencing as losing the sequence information in noise when used to detect ...
2
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1answer
34 views

Retrovirus Production

I have been having difficulties with low transduction efficiencies of my retrovirus production. I expand my plasmid of interest (on MiG-GFP plasmid) in DH5α E Coli for ~24 hours, purify with Qiagen ...
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1answer
75 views

Why does gaining a hydrogen in biology considered reducing a molecule? [closed]

I've learnt in chemistry that gaining electrons means reduction, while losing electrons means oxidation. But why is it in Biology textbooks I sometimes come across the term gaining hydrogen??
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249 views

Why people fear GMOs? Can't we map a plant composition?

My main question is can we map what a fruit is made of? For instance apples are made of 0.0002% of protein X, 0.00001 of protein Y, 0.001% of amino acid Z... etc... If we can, then my next question ...
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26 views

Concentration of degenerate primers should you dilute to?

I'm a little embarrassed to ask but when you have for example four degenerate primers and the end protocol says that the final primer concentration should be 10 µM working stock, should you make the ...
2
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1answer
44 views

How are lesions in the RNA corrected?

I quite understand why thymine is present in DNA. So we can mark it out where cytosine undergoes a reaction and is converted to uracil. Then we can repair the DNA. But how can we make that out in RNA ...
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29 views

Gene Sequencing and Plasmapper

Is there anything similar to this in Java (especially the circular map sequencing along with hover effect)? For information I would like to convey that I am using Plasmapper and BioJava for achieving ...
2
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52 views

How do C. elegans manage nutrition?

If there is ample amount of food, do C. elegans worms know when to stop eating or do they store extra energy? Could they put this extra energy to use by moving faster or putting more eggs?
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196 views

Difference between sequential and conformational epitope

Is the difference only in its structure? Like conformational epitope has 3D structure while sequential has a linear structure?(I have not studied biology since last 8 years and now I am going through ...
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29 views

Transcription rate expressed in microarray per hour

This article gives measurement of transcription rate and the unit they're using is microarray per hour. For example, at 27°C the average expression of their genes is 236.1 microarray per hour (page ...
2
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1answer
251 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 ...
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35 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 ...
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749 views

What is Pan for in pan-caspase?

A simple question (I could not find it on internet): What is Pan for in pan-caspase? Is it any different from the term 'caspase' ?
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61 views

In genome research, what is the problem in Mapping that may be caused by reads being too short?

In the following scenario: You were given short sequence reads of plant RNA obtained from a next-generation sequencing machine (fragments of 20–30 nucleotides in length). You attempt to map them back ...
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147 views

Role of isopropanol in plasmid isolation

In plasmid isolation, isopropanol is used to renature only the plasmid macromolecules after we precipitate all macromolecules(DNA, RNA, plasmids) using sodium acetate solution. How does it do that? ...
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181 views

Troubleshooting for Agarose Gel Electrophoresis

SI ran my 2% agarose gel on constant voltage (100 V). For a few times it worked at CV but, after a week or so the machine would automatically switch to constant current by itself. And this only ...
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294 views

How can Southern blotting be used to establish gene copy number?

I am reading a paper on a certain marine microorganism that can eat alkanes and therefore might be useful for cleaning up oil spills, etc. This sentence struck me as odd (although I am sure that the ...
2
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1answer
732 views

Orientations of codon and anticodon

So 5'-AUG-3' is the start codon on an mRNA transcript Does that mean that the tRNA anticodon is 5'-CAU-3'? Here, I see that the tRNA anticodon for methionine is UAC, and the chart doesn't specify ...
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1answer
976 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 ...
2
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2answers
192 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 ?
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1k views

Why is allolactose the LacI inducer?

For what reason(s) is allolactose, instead of lactose, the "natural" inducer of lac operon repressor?
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220 views

Can Taq polymerase be used instead of polymerase Vent exo (-)?

Instead of using polymerase Vent exo (-), can I go with the usual Taq polymerase? Do the PCR conditions change (the temperature and master mix concentrations ) in the these two conditions? Do the ...
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57 views

How to isolate host cell RNA (tRNA,mRNA,rRNA) from viral RNA?

A retrovirus with RNA genome infected a host cell. You would like to isolate the host cell’s RNA (rRNA, tRNA, and mRNA) from the virus RNA. What properties can you rely on to determine the three types ...
2
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38 views

Why is the resting potential of a neuron so close to the equilibrium potential of K⁺?

I know this has something to do with the K+ leak channel. I just don't understand how. I know that 3 Na+ are pumped out for every 2 K+ pumped in. This makes the cell interior net negative. I know ...
2
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1answer
29 views

Repair of cloning vector nicks digested with antarctic phosphatase and ligated with T4 enzyme

The vector product obtained by ligation between a vector previously digested with antarctic phosphatase lacks 5' phosphate groups. T4 ligase can ligate it to an insert with complementary sticky ends ...
2
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1answer
26 views

Separation of closely-sized isoforms

I have to separate two proteins of 86kDa and 80kDa respectively, however, I just cannot get a decent separation even in 6% polyacrylamide gel. To make matters worse, these two proteins are isoforms ...
2
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1answer
170 views

Active & passive transport question

If an element, ion or molecule is found in a cell is it possible to tell which method of transport was used? for example if a hydrogen or sodium ion was found in the cell could you tell if it got ...
2
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1answer
40 views

Kinesin-5 / cytoplasmic dynein spatial density distribution in neurons

Is there some way to experimentally determine the density distribution of Kinesin and Dynein in a Neuron? Fluorescence labeling would be impossible(?) as GFP markers would probably alter the motor ...
2
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1answer
3k 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 ...
2
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1answer
34 views

Transport of mammalian insulin in vivo

What is the mechanism for transport of insulin in mammalian cells?
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103 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
119 views

Transition/Transversion ratio

I'm taking a bioinformatics class and at this point we are just going over basic stuff about molecular biology. Half of the class is doing reading/research on our own, so there isn't much class time ...
2
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2answers
79 views

Molecular simulations without capping n- and c-terminus of polypeptide

When running molecular simulations, what is the consequence of not capping the n- and c-terminus? I have been following gromacs tutorials and haven't come across any that deal with capping the ...
2
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1answer
80 views

Determining sequence of oligoribonucleotide

Oligoribonucleotide X was treated with phosphatase (for removal of 3' and 5' - terminal phosphates), then with RNAase T1, which cleaves all phosphodiester bonds located in a 3' position of ...
2
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2answers
125 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 ...
2
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1answer
341 views

How to calculate or know by experiment the entropy of enzymes or protein?

How do you calculate or experimentally determine the entropy of enzymes or protein? In particular, I am interested in Boltzmann and conformational entropy, and Gibbs free energy. Any references are ...