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

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-3
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0answers
18 views

How is the protein removed from the DNA preparation? [on hold]

to determine how much DNA you have, you measure the absorption of the DNA.
4
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2answers
1k 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 ...
3
votes
1answer
97 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 ...
8
votes
1answer
63 views

How to validate the regulatory interactions inferred from gene expression data?

My algorithm learns regulatory interaction between genes using Bayesian Network approach from gene expression data. After the algorithm has converged to a network of interacting genes, how to validate ...
5
votes
1answer
562 views

What is the definition of a stringent/relaxed plasmid?

I have found a publication which proposes some definitions, including a definition for strict and relaxed replication. The definitions are: Relaxed control of plasmid replication. Relaxed control ...
4
votes
1answer
31 views

Western blotting with multiple antibodies

Normally I wash/detect with one primary/secondary-HRP antibody pair, strip, then wash/detect with the other primary/secondary-HRP pair which works well. However, I recently started working with a ...
-2
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0answers
24 views

what makes different materials to have different tastes? [on hold]

what makes different materials to have different tastes? For example: what we know as different colors is due to different frequencies of light and what we know as sound is due to the different ...
-4
votes
0answers
40 views

What are the 5 most expensive machines that can be found in a bio-lab? [on hold]

What are the 5 most expensive machines that are specific to a biology laboratory? In this case the type of biology laboratory that specialises in molecular biology and bioinformatics (study of cell ...
0
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0answers
28 views

coevolution by physical interaction between RNA sequences [closed]

There are several items that relate to the concerted evolution between pairs of sequences (especially structural) rRNA as a consequence of physical interaction in tertiary, or even quaternary ...
8
votes
2answers
355 views

More variation in proteins than genes. Why?

The Genome of a cell or organism is the same as that of the entire organism. However, the proteome of an organism is much greater than that of each cell (unless the organism is unicellular). How do ...
2
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0answers
12 views

Resource for finding Repressive/Inhibitory factors for a given gene?

I have a list of genes for each of which I'd like to find: A list of transcription factors that up-regulate the gene A list of inhibitory factors that down-regulate it. I used this tool on ...
5
votes
3answers
512 views

Should I dilute DNA with water or elution buffer?

I've extracted DNA using a kit and final step is eluted with buffer. If I must dilute my DNA samples, do I use the same elution buffer or can I use milliq water?
0
votes
1answer
13 views

Rosetta strain with chaperones for protein expression?

I am trying to purify a protein, and I was wondering if it is reasonably straightforward to obtain E.coli cells containing: -pGroe plasmids expressing chaperones. -Rosetta plasmids with codons that ...
3
votes
1answer
22 views

What's up with all the vague protocols? [closed]

I have lost count of how many protocols I've seen, including those supposed to be professionally written (such as manuals that come with kits from well known brands, or methods sections of papers in ...
1
vote
1answer
29 views

PCR directly on DNA binds to Ampure XP

During the library preparation it's possible to conserved Ampure beads during the different enzymatic reaction (it's the with beads strategy for improving yield, Fisher et al. Genome Biology 2011). ...
1
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0answers
14 views

how to get truseq's gel size selection when preparing libraries for exome enrichment?

i would normally use a 6 minute fragmentation as quoted by agilent but this gives post exome enrichment libraries of around 300bp or slightly over. Truseq want libraries to be 100bp larger. i am ...
2
votes
1answer
24 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 ...
3
votes
2answers
253 views

Why does methylation not occur in viral DNA?

Why does methylation not occur in viral DNA? Can viral DNA undergo the process of methylation? If not then why does this process does not occur in viruses?
1
vote
1answer
33 views

Degenerate primer design for DIG in situ hybridization

New to molecular and have learned to design primers from google/youtube so any info would help Would someone be willing to share their protocol for degenerate primer design? Breakdown: Trying to ...
4
votes
1answer
31 views

A few questions regarding immunology [closed]

I know that there is a variable region on antibodies which can recognize a wide variety of antigens, and that germinal centers create more "fit" antibodies to respond to an infection. So I was just ...
1
vote
1answer
38 views

Low copy numbers of plasmids

I have a plasmid with the P15A origin which apparently has a low copy number (see here). This would explain why my purification yeilds for subsequent digestion are low (gel shows the plasmid after ...
0
votes
2answers
56 views

Purifying a linear plasmid after restriction digest?

I expressed a yeast vector in E.coli and purified about 13µg of it. I then linearized it using a restriction enzyme, and attempted to gel purify it. I attempted this twice. The gel showed a clear ...
3
votes
1answer
51 views

PSI-BLAST website algorithm parameters

http://blast.ncbi.nlm.nih.gov/Blast.cgi In this website, when I want to apply the psi-blast algorithm on a sequence, under the section of algorithm parameters , what does PSI-BLAST threshold mean? ...
2
votes
1answer
35 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
votes
1answer
164 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: ...
1
vote
1answer
96 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
1
vote
1answer
41 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? ...
2
votes
1answer
99 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? ...
4
votes
1answer
52 views

Restriction Mapping - Homework question

I have trouble in solving this exercise. Exercise A circular plasmid of 10,000 base pairs (bp) is digested with two restriction enzymes,A and B, to produce a 3000 bp and a 2000 bp bands when ...
6
votes
2answers
90 views

Can an organism process H₂O into H₂O₂?

In an answer to a recent question on Worldbuilding, I suggested that an organism convert $H_2O$ into $H_2O_2$. I suggested a few processes that yielded the desired final result ($2H_2O \rightarrow ...
1
vote
0answers
56 views

Solid phase use in HIV rapid tests [closed]

I have a 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 region of ...
4
votes
1answer
76 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
votes
1answer
42 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 ...
2
votes
1answer
77 views

How does temperature influence the rate of protein degradation?

For computer modeling purposes, I am looking for some referenced quantitative measurements of the effect(s) of temperature on biochemical reactions. Question In particular, my question is: How does ...
1
vote
2answers
28 views

C.elegans and antioxidants

How would you test the effect of antioxidants on C.elegans lifespan? Is this done through feeding E.coli with antioxidants and then C.elegans with E.coli, or is there another method?
0
votes
1answer
43 views

Apoptosis vs necroptosis

I understand that apoptosis and necroptosis share the same upper part of the pathway, but I cannot seem to distinguish when is each one activated? From my readings, it seems that when procaspases 8 or ...
1
vote
2answers
143 views

Book Recommendations: GRE Subject Test In Biochemistry, Cell And Molecular Biology

There are probably a lot of really good answers that may vary significantly in terms of content. I'm looking for a set of books that I can read in preparation for the GRE Subject Test In ...
2
votes
1answer
76 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?
2
votes
1answer
24 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 ...
6
votes
1answer
81 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 ...
5
votes
1answer
80 views

enzymes that stabilize DNA loops

As a follow-up of a previous question, I would like to know what enzymes or protein complexes have been used to manipulate DNA samples into stabilizing DNA loops. I have read that cohesin is one of ...
2
votes
1answer
48 views

How small does a nanobot have to be to “swim through the brain” and access any neuron it wants to?

I read on this question What is in the space between neurons in a brain? that there is actually not much empty space in a brain. But my question is slightly different. Is there a visual demonstration ...
1
vote
1answer
17 views

Centriole genes Knock-out Experiment in Common experimental animals?

Anyone know of any experiments that have knocked out the genes for producing centrioles in a worm, mouse, fish, fly or whatever animal? Are the genes for centrioles even identified? It has been shown ...
7
votes
1answer
173 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 ...
5
votes
2answers
227 views

poor RNA quality from zebrafish embryos

Does anyone routinely do RNA isolation from zebrafish embryos? I have embryos from different stages but all below 24hpf. This is the protocol I follow: Take 10-20 embryos Wash once with milliQ ...
1
vote
1answer
51 views

How to separate peptide on SDS PAGE?

I want to separate 1600 Da peptide on SDS PAGE, on 20% GEL I didn't get any band, How can I separate on PAGE? Can I use 25% SDS PAGE?
1
vote
2answers
95 views

Photosynthesis: Splitting Water

The splitting of water is an endergonic (non-spontaneous) reaction, and thus would require energy (chemical work to be done) in order to happen. In Photosystem II, an enzyme catalyzes this splitting, ...
9
votes
1answer
113 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 (Big genome size)? energy for copying? raw material for copying? space in the cell? Maintenance cost (matter and ...
3
votes
2answers
35 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 ...