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

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103 views

Suggestions for an experiment? [closed]

I would appreciate suggestions for my year-long AP Biology project. Such a project would consists of a standard experiment, to be done over the period of a year, so it would have >to be fairly ...
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2answers
53 views

The role of antibodies interacting with cancer

I'm learning about antibodies. As I understand it, antibodies detect stranger cells/bacterial/viruses by the molecules present in their membranes. In cancer cells, the cancer cell have produce some ...
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1answer
82 views

Preparation of normal DNA polymerase

i don't need Taq polymerase. Just need normal DNA polymerase. So the main idea is preparation polymerase directly from Lactobacillus without using cloned vector. My idea is electrophoresis using ...
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1answer
81 views

What in soy are people allergic to and will these allergens be passed along if an animal that has eaten soy is consumed?

I have two related questions about soy allergies and possible reactions. What is it that are humans who have soy allergies actually allergic to? What is the allergen? If an animal eats soy, are ...
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1answer
45 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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1answer
38 views

In vitro transcription, contamination problem

I am using a RNA which is in vitro transcripted before I started my project. It turned out it is not prepared properly and has DNA contamination. Instead of perform the in vitro transcription again, ...
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1answer
41 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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39 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, ...
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1answer
46 views

Clarification of the procedures to amplify cDNA needed

The typical procedure to obtain cDNA from genomic extracted DNA is the following: RNA is extracted from desired tissue, RNA/DNA hybrid is obtained by virtue of reverse transcriptase RNA dependent DNA ...
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2answers
55 views

How does promoter sequence affect initiation?

I don't know if this might have been highlighted in recent research, but a textbook I have states that "the exact way in which promoter sequence affects [transcription] initiation is unclear" I'm ...
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2answers
265 views

Gel electrophoresis question

Leena is a molecular biology student. She purifies two fragments of DNA, 800 and 300 base pairs long. These were obtained from a plasmid after digesting it with HindIII. Each of these fragments has a ...
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1answer
78 views

how to clone a gene fragment in two direction as sense and antisense in vector

I have a sequence ATG GGG CCC TTT AAA TAA and want to use it as antisense RNA in my vector. How should I clone it? I am confused with the direction of my clone. I looking for the direction and ...
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1answer
53 views

DNA preparation for sequencing

In shotgun sequencing method or some related method that DNA is break up into random fragment. The fragment that have about 3kb in size is inserted into plasmid by enzyme ligase and then plasmid ...
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280 views

How do viroids propagate to other hosts?

As viroids are simply non-encapsidated non-coding RNA molecules, it is difficult to imagine a mechanism for their continual infection, other than horizontal transfer which doesn't seem to be the case. ...
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1answer
84 views

Hardy-Weinberg applied to three alleles and stimation of allele frequencies

I have this equation: Corresponds to HW in equilibria with three alleles: $(p+q+r)^2=1$ Expanding the square results: $p^2+2pq+r^2+2pr+q^2+2qr = 1$ I need to separate homozygous and ...
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1answer
30 views

Amount of reverse transcriptase in µg or mM for qRT-PCR

I am trying to calculate a titration amount for a molecule which I would like to use in my PCR samples. Different molecules have different densities so I would like to calculate the appropriate ...
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1answer
25 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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1answer
41 views

Missing 4$H_2O$s (per glucose) in Cellular Respiration… Where can they be?

I having trouble understanding the equation of the cellular respiration. The thing that bothers me is the number of $H_2O$ molecules. Generally, cellular respiration is written thus : $C_6H_{12}O_6 + ...
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1answer
42 views

How to relate human metabolic model reactions and cancer drug targets / reactome items?

Trying to find a way to take a cancer drug (from CancerDR, for instance) and infer the metabolic reactions that are affected by it in the Human Metabolic Model. Essentially, I would like to know ...
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1answer
52 views

Why bacteria produce light?

After seeing this video I am curious to know why the free living bacteria produce light. What advantage will they have ? Or is it just an "unintentional" result of one of their pathways ?
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21 views

Vesicular and non vesicular transport

I have to classify them either as transported in vesicles or without vesicles. What I think - Non-vesicular 2.vesicular 3.vesicular 4.vesicular 5.non-vesicular 6.vesicular 7.vesicular ...
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1answer
62 views

Shine-Dalgarno sequence and expressing proteins

Shine-Dalgarno sequence present in the prokaryotic mRNA plays a role in initiation of translation. In eukaryotes a Shine-Dalgarno like sequence is present but does not play an important role in ...
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1answer
41 views

Protein Structure Statistics [closed]

Could you show me references of any works you know of where, 1- they take all the known protein structure data (that is the coordinate data of all the proteins in PDB database lets say or elsewhere) ...
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1answer
69 views

At what temperature does DNA re-anneal?

If primers weren't present in PCR at what temperature would DNA re-anneal? I am wondering how primers manage to bind on DNA strands before DNA manages to anneal back together.
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2answers
39 views

Why don't raisins lose water when in air?

It is common that we soak raisins in water, and they acquire water by endosmosis. That means that the cells are permeable to water, right? Then why don't they lose water when kept in air? Or kept in ...
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1answer
60 views

Complementarity of cDNA

Strictly speaking, what is the definition of cDNA? This confuses me, since usually it is said to refer to DNA that is complementary to mRNA. Is this correct? Is it restricted to mature mRNA? I ...
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1answer
153 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 ...
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73 views

Do mitochondrial DNA have repair enzymes?

Mitochondria are considered endosymbionts originated from a prokaryote. So, do they have a DNA repair mechanism ?
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48 views

Isoprenoid Diphosphate Concentration in Yeast saccharomyces cerevisiae

Isoprenoid Diphosphate(IPP) is an important metabolites which is precursor in lot of secondary metabolites like Dolichol diphosphate, ubiquinone, prenylated proteins and carotenoid (not synthesized ...
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1answer
39 views

In vitro enzyme production

I need to express a protein in vitro but I don't know where to start. I will likely do a T7 transcription protocol but for translation I am not sure what to do. Are there any good kits?
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1answer
234 views

What is solid-phase bridge amplification?

During Illumina sequencing there is a step called bridge amplification by which DNA is amplified by isothermal enzymes. What is this stage, and how does it work?
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1answer
172 views

Why does the hydrolysis of ATP increasing entropy increase the Phosphoryl-­‐transfer Potential?

I am currently taking a biology class. I do not understand this concept. I understand that the electrostatic repulsion of the negative charges, resonance stabilization and hydration stabilization all ...
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1answer
440 views

Molecular cause of cramps, spasms and strengthening in muscles? (incl. intro to muscle contraction)

When motor neurons are stimulated to trigger an action potential, this potential propagates down the spine, eventually reaching a neuromuscular junction, causing the release of acetylcholine (ACh). ...
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85 views

Which mutagen is responsible for pancreatic cancer [closed]

Which mutagen, aflatoxin or acridine orange (an intercalating dye) would most likely lead to an increased incidence of pancreatic cancer. Why?
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0answers
41 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 ...
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35 views

How is the number of histones being related to the number of chromosomes [closed]

Please name me at least one reason. I am referring to rats and to humans, as well.
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467 views

Bacterial 16S rRNA PCR amplification with universal primers

I am doing an experiment to amplify the 16S rRNA gene from bacteria present in gut contents of fish. After extracting DNA, I perform one-step PCR with universal bacterial primers (27F, 1492R) and I ...
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2answers
98 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 ...
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0answers
35 views

Can DNA ploidy analysis differentiate field cancerization effects of oral potentially malignant disorders?

Can DNA ploidy analysis differentiate field cancerization effects of oral potentially malignant disorders [ Leukoplakia, erythroplakia, oral submucous fibrosis, etc] ?
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37 views

Protocol for immobilize oligo to solid

I'm learning about microarrays and understand that an oligonucleotide can be immobilized to a solid like a CD acrylic plastic by modifying the oligo. Does anyone have a good protocol for modifying a ...
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0answers
34 views

Are stable proteins tagged by ubiquitin?

What would happen if an extremely stable protein was created because of some error? Would that protein be tagged by ubiquitin?
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0answers
440 views

Low RNA yield, low 260/230 ratio

I extracted total RNA from animal tissue using the Qiagen RNeasy kit, however my RNA yield was extremely low and the 260/230 ratio was around 0.3. This is the protocol I followed: The animal ...
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0answers
104 views

What mammalian cell lines are easiest to grow and transfect?

I'm working with HEK293 cells, which are very easy to transfect. Does anyone have other suggestions for cell lines with very high transfection efficiency? Ideally the chromosomes would also be diploid ...
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0answers
44 views

Experience with Detox Cloning

I just saw this paper. A Highly Efficient Molecular Cloning Platform that Utilises a Small Bacterial Toxin Gene I was curious if anyone had experience with this cloning strategy using the IbsC ...
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0answers
47 views

Solid phase use in HIV rapid tests

I have another 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 ...
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3answers
105 views

Pump to separate protein/DNA instead of electrophoresis?

I know that DNA molecules or proteins can be separated in electrophoresis because the electrical charge is used to pull the DNA through the gel. So instead of using electrical, can I using physical ...
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2answers
80 views

How to determine the actual flux in the human metabolic model of a cell line?

The problem to be solved is to determine what the flux values are for the different reactions in the human metabolic model. As far as I understand, a good way to do that would be to use gene ...
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1answer
115 views

Why not use SDS-PAGE as a method to detect viruses?

Recently, I have been researching about DNA and I know the most popular method for detecting viruses is based on DNA. After learning about proteins, I wonder why we do not detect viruses based on ...
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2answers
69 views

Problem on finding sequence of Taq polymerase

As i know, Taq Polymerase can be found in Thermus aquaticus, so i do a search for protein list of Thermus aquaticus and have this : http://www.ncbi.nlm.nih.gov/genome/proteins/1724?project_id=55053 . ...
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1answer
36 views

Identifying mutations in a yeast strain

Cells of a triple-mutant yeast strain (leu- his- trp-) were spread either on minimal medium or on minimal medium supplemented with various combinations of histidine, leucine, or tryptophan. The ...