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

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3
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0answers
49 views

E. coli not growing in liquid medium

We are regularly doing bacterial transformation and subculture from plate to liquid media to extract DNA. This usually goes very well and is straightforward, but occasionally, the colonies that grew ...
2
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3answers
31 views

Looking for good reference book in molecular biology [on hold]

I am starting to work in molecular biology/ molecular genetic and I am looking for a really good book containing the main concepts and mostly the more recent techniques. I would be interested in a ...
1
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1answer
40 views

Is the amount of dNTPs rate limiting for very long PCR products?

I'm using the Q5 system and I'm PCRing a product that will have a final length of around 9500 bases. I have noticed that the product is there but very faint. The primer seems to be used up. So my ...
4
votes
2answers
84 views

What are the “minimum requirements” for a single cell?

I saw a description of the "minimum requirements" for a cell at http://creation.com/origin-of-life in the section called "What are the minimum requirements for a cell to live?" and I'm wondering if ...
4
votes
1answer
48 views

Does suffocation pain involve nociceptors?

I never suffocated myself so not entirely sure, but when you suffocate, it's painful, right? But of course different kind of pain than being injured or sick. What I'm wondering is, if the "painful" ...
2
votes
1answer
20 views

Why does the inhibition of translation initiation cause the accumulation of 80S ribosomal monosomes?

As I have read in (1), inhibition of translation initiation will increase the number of 80S ribosomes while decreasing the fraction of polysomes due to the polysome-runoff. The net effect is to ...
2
votes
1answer
20 views

What is the protein sequence taken as input in the Path-A prediction system

Considering the Path-A based metabolic pathway prediction (http://nar.oxfordjournals.org/content/34/suppl_2/W714.short). It uses machine learning for pathway prediction. Suppose that the input was a ...
6
votes
1answer
217 views

How many RNA-binding proteins can simultaneously bind on a single mRNA?

Typically, how many RNA-binding proteins can simultaneously bind to a single mRNA? Or said differently, how many "binding sites" does an mRNA have? What order of magnitude? I am interested in RNA ...
1
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0answers
43 views

Recommend any Molecular lab LIMS (Laboratory Information Management System)

We are looking to develop or customize a LIMS for our molecular lab. Do you know of a LIMS that you've used in a molecular lab before, or one that could be used. Thanks for the help (Edit) Some of ...
7
votes
2answers
194 views

What is the difference between sequence, reads, and contigs of genetic material?

Can someone explain the differences between sequence, reads, and contigs of genetic material such as DNA, if possible with an example? I am new to bioinformatics, and I have not found any conclusive ...
11
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2answers
76 views

How can we predict protein folding in silico?

Currently, there is a lot of research focused on solving the folding patterns of proteins using computers (Folding@Home, https://fold.it/portal/, etc.). The question that I have is: How do you know ...
0
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1answer
22 views

How is cancer associated with host pathogen interaction?

Does cancer by any chance fall under host pathogen interaction domain? What I mean to ask is that, is there pathogen interaction involved in cancer? I went through this article: ...
2
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0answers
15 views

Does blood typing still provide a use for ancient tissue analysis?

Modern techniques. In recent years, DNA sequencing has become extremely cheap. This, compounded by the ability to PCR miniscule samples to viable samples for analysis, means that aDNA can be ...
2
votes
2answers
31 views

How can 3' end polyadenylation protect cellular RNA from ribonuclease degridation?

In this link, it states: It is worth noting that all intracellular RNAs are protected from RNase activity by a number of strategies including 5' end capping, 3' end polyadenylation, and folding ...
7
votes
1answer
60 views

Double stranded nucleic acids are more 'durable' than single stranded nucleic acids?

I'm struggling with a question I've been asked. "Why is double stranded genetic material more 'durable' than single stranded one?" I know that double stranded genetic material is more stable due to ...
0
votes
1answer
25 views

What is meant by clonal isolates?

With respect to this website: http://horfdb.dfci.harvard.edu/ what does the term clonal isolates mean?
2
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0answers
21 views

Storing Taqman plates with polymerase in fridge before running

I'm going to be running some Taqman assays on cDNA generated from RNA collected from various samples, and will end up running a bunch of plates (right now the setup is 1/2 a plate per sample, ...
2
votes
1answer
25 views

Which is the guideline to choose a molecular target to identify vertebrate hosts from arthropod bloodmeals?

There are some molecular targets to identify vertebrate hosts from arthropod bloodmeals including the Cyt b gene and the COI gene. Which are the standards or characteristics that I have to bear in ...
1
vote
1answer
21 views

specific numbers of neucleotides in okazaki fragments

Why there is specific number of nucleotides in okazaki fragments...? Okazaki fragments are form in lagging strand during the process of DNA replication.
8
votes
2answers
26k views

What is the function of the RNA primer in DNA replication?

During DNA replication, RNA primase puts an RNA primer in the lagging strand. What is the function of this RNA primer? Why can't the enzymes put DNA fragments directly?
3
votes
2answers
53 views

What physical force attracts the anti-codon on tRNA to the codon on mRNA during translation?

What physical force attracts the anti-codon on tRNA to the codon on mRNA during translation? I know that these two bond together, but what actually makes the tRNA move through the cytoplasm to the ...
3
votes
2answers
37 views

Macrophages and what they consume

Do macrophages in the dermis consume any foreign invader to the body, or simply pathogens and cellular debris? For example, would macrophages consume vitamins or medications injected into the dermal ...
1
vote
0answers
26 views

What will happen to transcription if histone acetyltransferases are removed from a eukaryotic cell?

If HATs are destroyed in a eukaryotic cell, how will gene expression level be affected? Will all gene expression be affected or just be slowed down and most gene will still be expressed?
0
votes
1answer
31 views

(Genetics) Is a silencer the same as gene silencing (heterochromatin)?

Is silencer the same as gene silencing? I know that gene silencing refers to those heterochromatin concentrated at the telomeres or centromere. It is also related to methylation. But what about a ...
4
votes
1answer
122 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 ...
10
votes
2answers
6k views

Why is PEG important for efficient yeast transformation?

One way to do an yeast transformation is by using lithium acetate, a single-stranded carrier DNA, and PEG (1). I was wondering why is the polyethylene glycol important for the efficient ...
0
votes
0answers
22 views

What is a “NuMA protein”?

I just got out of a lecture on mitosis and this part was incredibly confusing. After searching my textbook it doesn't cover it that well either. The professor mentioned a NUMA protein that is ...
1
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0answers
36 views

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 ...
1
vote
2answers
73 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?
2
votes
1answer
96 views

Enzyme Assay - pectinase

During assaying an enzyme at high temperature, the substrate (Pectin) is degraded by the high temperature rather than by enzyme, so, how can I minimize degradation of the substrate by the temperature? ...
3
votes
2answers
55 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 ...
5
votes
1answer
56 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
votes
1answer
39 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 ...
1
vote
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 ...
2
votes
1answer
272 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 ...
5
votes
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, ...
4
votes
1answer
36 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 ...
1
vote
1answer
32 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?
1
vote
0answers
12 views

OCT-1 transport in Enterocytes

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 ...
3
votes
1answer
29 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 ...
10
votes
2answers
93 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 ...
5
votes
0answers
52 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 ...
4
votes
2answers
52 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
votes
1answer
64 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
votes
1answer
36 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 ...
0
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0answers
20 views

Workspace preparation and cleanup for DNA work

What steps should be taken in a molecular lab environment to help ensure that DNA samples/stocks are not contaminated, or contaminate other objects in the lab?
4
votes
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?
1
vote
0answers
19 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 ...
1
vote
2answers
27 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
votes
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 ...