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

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Importance of organic chemistry in biochemistry/molecular biology? [closed]

I'm halfway through my biochemistry and molecular biology degree in Australia and i'm concerned about a lack of organic chemistry. My friends studying biochemistry back home in Canada take units upon ...
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2answers
84 views

What is the importance of urea in mass spectrometry?

What is the importance of urea in mass spectrometry? We use 8M urea to FASP our proteins prior to mass spectrometry. What is the significance of using 8M urea? and how does it affect the proteins?
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2answers
60 views

Need help in codon optimization

I want to chemically synthesize a GFP gene for expressing in rice.I am using the IDT Codon Optimization program and found that not all the codons are 100% optimized. Some of them are the 2nd or 3rd in ...
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1answer
62 views

Is it possible to express a mutant gene only in a specific tissue?

Imagine that someone tries to develop a knockout mouse for a gene, but this result in lethality for the homozygous. Is it possible to express that mutant gene only in a specific tissue of interest to ...
2
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1answer
42 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 ...
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1answer
25 views

Reliability of Sanger sequencing

Hailed as the "gold standard" of sequencing, I was wondering to what Sanger sequencing owes its incredible accuracy to. If possible, I would like a quantification of the accuracy (because I doubt it ...
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1answer
300 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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1answer
48 views

Do non-enzyme catalysed reaction pathways exist?

Can their be a kind of chemical reaction pathway in a cell, that is catalyzed or regulated but NOT necessarily by enzymes? I could not find anything on Google. I have almost no background in biology, ...
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1answer
47 views

blue/white screening - results are opposite as expected

We inserted GFP-gene into plasmid Bluescript SK+, transformed E. coli with this construct, and then plated on an agar plate with Ampicillin and X-gal to do a blue/white screening. We got blue colonies ...
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1answer
34 views

What are the factors on which protein-protein interaction depend? [closed]

Does protein-protein interaction only happens when one of them is basic and the other acidic? Do protein interactions also depend on the protein structure? Are there more factors?
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0answers
12 views

OCT-1 transport in Enterocytes [closed]

Does Oct-1 transport lactate or any of its products and in which direction (intracellularly to extracellular, the opposite, or inter-nuclear). The literature talks about Oct-1's role in neuronal ...
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1answer
37 views

DNA length and annealing kinetics

I have a mixture plasmids and undesired short linear fragments that share the same sequences. During denaturation and annealing, I would like the plasmids to 'find each other' before annealing to the ...
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2answers
99 views

Do macrophages get nutritional value from the pathogens that they eat?

X-posted on reddit AskScience here. I know that macrophages engulf foreign bodies and transport them to various waste excretion pathways (sorry if the terminology is wrong), and if the foreign bodies ...
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63 views

SDS Laemmli Gel that did not solidify

In my lab class this week my partner and I were making a SDS-PAGE Laemmli Gel. We made the resolving gel solution which was made of 30% Acrylamide, DI water, 1.5 M Tris-HCL pH8.8, and 20% SDS. We ...
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2answers
56 views

How can (or did) Deinococcus radiodurans continue to evolve after developing resistance to mutation?

Deinococcus radiodurans has a remarkable ability to resist damage to its DNA due to radiation, dehydration or (to my knowledge) any other source. It keeps multiple copies of its genome and has a ...
2
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1answer
75 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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1answer
37 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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2answers
32 views

Transformation efficiency

I accidentally spun down the cells after heat shock treatment before adding the media into the tubes while doing transformation? will the transformation work?
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0answers
20 views

Question about cytoskeleton coordination

I am trying to study for a biology and I am having some confusion over the following topic. Can anyone help explain/ shed some light on the concepts of Rho family GTPases. Is it true that we have Rho ...
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2answers
31 views

How does the placement of it's motor domain affect the directionality of a motor protein?

I'm going over some of my notes, and I have written down that motor proteins with + cytoskeletal directionality have a reversed schematic representation relative to - directed motor proteins. Then I ...
2
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2answers
56 views

What is meant by nucleated in this context?

Hi I am little confused in one of my classes; because the teacher will say that " the actin is nucleated by the ARP 2/3 complex" or "microtubles are nucleated by the centrosome". Unless I am ...
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1answer
26 views

Why introduction of an extra copy of a gene related to pigmentation causes RNA interference in Petunia?

RNAi became famous after Fire and Mello experiment in C.elegans; however, it had been observed before. In the 80's, Jorgensen was trying to increase pigmentation of Petunia flowers by introducing ...
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2answers
73 views

Why do mitochondria fuse together?

Contrary to all of the textbook images of mitochondria that I have seen over the years, I had just learned that the mitochondria within a cell form a dynamic branching network along microtubule ...
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4answers
2k 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 ...
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1answer
57 views

Is it possible that a set of functionally related proteins in a pathway fulfill different functions? [closed]

Could it be that a given pathway of enzymes (or proteins in general) may fulfill different purposes in a cell by for shifting partners? Say protein A activates B, B activates C and C has a specific ...
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1answer
63 views

MHC restricted peptide

What is an MHC restricted peptide? I got this definition from wikipaedia, but cannot exactly extract what the phrase MHC restricted peptide means. MHC-restricted antigen recognition, or MHC ...
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0answers
44 views

CRISPR Knock in

Using the CRISPR/Cas9 technology, it is possible that after inducing a DSB with the Cas9 endonuclease guided with an RNA designed by the user and using a template DNA, get a desired Knock-In (KI) by ...
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2answers
535 views

How can CI repressor both activate and repress $P_{RM}$ promoter found in $\lambda$ phage?

I'm reading a paper where the authors constructed a toggle switch that uses bidirectional $P_R/P_{RM}$ promoter found in the $\lambda$ phage. There are 3 binding sites - $O_{R1}$, $O_{R2}$, $O_{R3}$ - ...
2
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1answer
51 views

DNA polymerase in PCR (polymerase chain reaction)

Can the DNA polymerase in PCR (polymerase chain reaction) recognize both DNA and RNA for use them as template? I want to know is it possible if my primers bind to an contaminant RNA and then any DNA ...
2
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0answers
24 views

Can encoding of miRNAs in introns not cause dysregulation of gene expression?

For example, consider a gene X which has an intron which encodes a miRNA that downregulates expression of another gene Y. When the gene X is translated, this miRNA will appear as a byproduct of ...
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0answers
26 views

Why can't some organisms match miRNA perfectly to the target mRNA like in plants? [closed]

What causes other organisms to be impaired in making perfect matches like plants do and is there a way to increase matching?
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1answer
28 views

What factors, other than its homochirality, make our “brand” of biology unique? [closed]

If humans were to discover organisms on another planet, it is supposed that (unless both we and they were seeded by the same source) we would have nothing to fear from alien pathogens, as they would ...
1
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1answer
58 views

Does DNA Ligase look for complementarity in sticky ends?

Can DNA ligase seal two non-complementary sticky ends if the reaction is incubated at the optimum conditions ? I do almost 100% double digestion in the lab. However, during ligation, when I do a ...
9
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1answer
148 views

RNA migrating slower than DNA on Formaldehyde Gel?

So I ran into an interesting problem. I'm getting a linear DNA band that is twice as long (4x bases, but as denatured probably only 2x) as an RNA band running at the same size in a formaldehyde gel. ...
4
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1answer
26 views

Oligonucleotide purification with desalting

I have ordered 36bp oligonucleotides that anneal to each other and create sticky ends to be cloned in a vector afterwards. I have tried cloning many times with different methods and I failed. Now I ...
4
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1answer
77 views

Nomenclature of genes and proteins

With respect to this paper: Identification of Host Proteins Required for HIV Infection Through a Functional Genomic Screen In the abstract, I found names such as Rab6 and Vps35 and TNPO3. So Rab6 ...
2
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1answer
25 views

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 ...
3
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2answers
45 views

Are restriction enzymes active at −20 °C?

I have digested my DNA with NotI enzyme and put it in the −20 °C freezer without heat inactivating it. Can restriction enzymes work at −20 °C? Should I expect STAR activity?
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2answers
50 views

Can molecular genetics make a boolean variable from a continuous variable?

In the same kind of idea than this question. Gene expression are regulated through complex interactions. The concentration of enhancers and repressors is an important aspect that dictate the level of ...
2
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1answer
103 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??
12
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1answer
1k views

How is Taq polymerase produced?

I've seen Taq polymerase being marketed as either "native" or "recombinant". I understand that the recombinant version is produced by specially modified Escherichia coli strains that have the gene for ...
7
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3answers
60 views

Mechanism by which $lacI^{d}$ is a dominant mutation, impairing the function of normal copies of the Lac Repressor

Jacob-Monod model for the lac Operon was based on experiments using two strands of bacteria which constitutively expressed $\beta$-gal: $I^{c}$(mutation in the gene lacI , which encodes the repressor) ...
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1answer
96 views

What are genome wide functional linkages?

There are two types of interaction classifications used to describe Protein-protein interactions, namely physical and functional. Whilst physical interactions are obvious in nature and methodological ...
3
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1answer
105 views

What happens to the precursor protein's signal sequence after it is cleaved?

Where does this signal sequence "go" after it has been cleaved by signal peptidase and what is its next function?
3
votes
1answer
101 views

What type of flask should I use to culture NTERA2 embryonic cancer stem cells?

I'm just starting my MSc research and I am in the process of making a list of equipments/consumables to order. Is there a specific flask in which I can culture NTERA2 (NTERA2/D1) cell line?
6
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1answer
99 views

Why do most organisms have negative supercoiled DNA?

It has been observed that in nature most organisms have negative supercoiled DNA and that few organisms have positive supercoiled DNA. Some of the organisms that have the positive supercoiled DNA live ...
1
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1answer
22 views

How to correlate the pattern by which CAP activator from E.coli binds to DNA and its mechanism of action?

The catabolite activator protein (CAP) activates the expression of more than 100 genes involved in secondary sugar metabolism in E.coli. Apparently, it always binds in sites that are away from -10 and ...
5
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1answer
212 views

Why is E.coli used as a model?

Is there a reason for the choice of E.coli as a model for many bacterial systems? Other bacteria such as B.subtilis are also used, but why is E. coli preferred?
7
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3answers
4k views

What are the advantages and disadvantages of using beta-galactosidase compared to luciferase as a reporter gene?

In the University labs, we have used Beta-galactosidase as a reporter gene to quantify the expression initiated by the stress-response promoter in yeast. This was done by exposing one of the two ...
5
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2answers
172 views

What is the IC50 exactly?

I am reading the paper "Activity of the bcr-abl kinase inhibitor PD180970" but I don't understand how IC50 works on table 1. Can you tell with simple words and give me an easy example?