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

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23 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? ...
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1answer
146 views

Importance of knowing GC Content of an organism

I was looking at the GC content percentages of few organisms. I also know calculating the GC content percentage. But, what I want to know is, what information would we get., let us suppose if human ...
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1answer
38 views

Complementarity Determining Regions (CDR)

Complementarity determining regions (CDRs) are part of the variable chains in immunoglobulins (antibodies) and T cell receptors, generated by B-cells and T-cells respectively, where these molecules ...
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1answer
22 views

Degenerate Alignment Analysis

Can someone please tell me what is Degenerate alignment analysis? Could not find a good article on the internet that could help me understand what it means? (I have not studied biology since last 8 ...
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1answer
20 views

Microarray probe and target

In a microarray, which one is called a target and which one is the probe? the one that is added later , is that the probe or the one present in the slots of the microarray, that is the probe? (I have ...
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22 views

TAA- Tumor associated antigen

An approach to find Tumor associated antigens is based on transfection of expression library made from cDNA into cells expressing desired MHC haplotypes. Can someone please explain what this line ...
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1answer
19 views

Why can't NMR be used on large RNA molecules?

While reading my Molecular Bio textbook, I came across a sentence that stated that NMR cannot be used on large RNA molecules to determine their structure. Why is this? Is it because RNA is single ...
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1answer
46 views

Protein Isolation from mammalian cells

What non-SDS buffer can I use to isolate protein from mammalian cells? The protein is intended for testing electron transport chain complexes.
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1answer
25 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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1answer
37 views

Localization of B and T cell

What does localization of B-Cell mean?? "Localization of B and T cell in allergens may not coincide". What does this statement mean? (I have not studied biology since last 8 years and now I am going ...
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1answer
27 views

What is the expected effect of pH on the activity of a fungal pectinase?

I am working on an enzyme assay for a fungal pectinase.I assayed the enzyme in different buffers from pH 1-12.5 However,the enzyme has good activities starting from pH1-10.5. Is it possible to have ...
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1answer
40 views

Propensity Score for amino acid

What is the meaning of propensity score of amino acid? How is it calculated? (I have not studied biology since last 8 years and now I am going through it because I need it for my research. So if ...
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20 views

Bipolar minus end directed molecular motor

like the title explains I am looking for a minus end directed bipolar molecular motor. Any ideas? Best wishes.
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1answer
21 views

How to identify the genes that distal enhancers pair?

I am writing a project proposal and I have to talk about this problem: how to identify the genes that distal enhancers pair? I am really new to this topic and I don't know what it is all about. I ...
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2answers
38 views

Western Blot: Fischer ECL substrate 2:1 instead of 1:1?

i recently developed a b-actin-ab-incubated PVDF-WB-Membrane with the Thermo Scientific Pierce ECL Kit. According to the manufacterers orders i mixed Reagent 1 and Reagent 2 1:1. But unfortunately i ...
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3answers
3k 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 ...
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1answer
69 views

What is a protective epitope?

What is a protective epitope? An epitope is basically a part of antigen. So does it mean that when the epitope combines with an antibody, it helps in the functioning of the antibody instead of going ...
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0answers
35 views

What does the term 'epitope mapping' mean? [closed]

Epitope mapping means identifying the binding site of antibodies on the target antigen. This means that the site to be identified is part of the antigen and not antibody, am I right?
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1answer
96 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? energy for copying? raw material for copying? space in the cell? Maintenance cost (matter and energy)? time to copy is ...
6
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2answers
206 views

Why does replication require primers while transcription does not?

In transcription, there is no need for any primer. I guess the basic mechanism of DNA polymerase & RNA polymerase is the same. So why does replication have the need for a primer?
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2answers
113 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 ?
2
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1answer
48 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
81 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
40 views

How do you express and detect GLP-1 receptors?

I am currently working with a peptide which is an analogue for glp-1, but during invitro studies am not able to detect for the presence of GLP1- receptors. The cell line used is Min-6. How do I detect ...
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1answer
45 views

What regulates the timing of the motion of molecular machines during DNA Replication?

This question is about this video I found on Youtube. I just want to know what is the mechanism which regulates the timing of motion of the parts of these molecular machines. I know that those big ...
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1answer
33 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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0answers
24k views

Does CPH4 exists? [closed]

I just see the film Lucy, it talks about a molecule called CPH4 which increase human capabilities. Of course it's a film and so it's not scientific facts. During an interview the producer says that ...
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1answer
41 views

Why do oncogenes show genetic dominance?

As we know that tumor suppressor gene causes cancer only when both the alleles are recessive in nature.But in case of oncogenes if only one allele is dominant it can cause cancer.Why in case of ...
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2answers
834 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 ...
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4answers
83 views

determine longest poly-purine tract

how to determine/ find the longest poly-purine tract in any genome and this needs to be on the E. coli genome . is it to figure out the polypurine tract and then figure out the longest chain ? or is ...
3
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1answer
63 views

Consumption of NAD+ in glycolysis

Out of 10 steps in glycolysis, only one reaction- Glyceraldehyde 3-phosphate (G3P) to 1,3-bisphosphoglycerate (PGP), uses NAD+ and thereby producing NADH. Furthermore, this very step is solely ...
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1answer
84 views

Selection on linked loci in a diploid population

Let’s consider two linked loci $A$ and $B$ that are both bi-allelic. In consequence, we have four different possible haplotypes $A_1B_1$, $A_1B_2$, $A_2B_1$, $A_2B_2$, which frequencies are $X_1$, ...
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3answers
1k 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 ...
3
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2answers
283 views

How are DNA segments selected in PCR?

I understand that in PCR we're able to amplify only selected portions of the DNA... however despite reading it from multiple sources, I cannot figure out how this selection actually takes place. I ...
2
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1answer
62 views

Why does the pET- expression vector contain a LacI gene additionally to the one in the genome?

The pET plasmid is used for protein expression with T7 promotor in expression strains, such as E.coli BL21(DE3) It contains a lacI gene which codes for the lac repressor protein, a protein of ...
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1answer
189 views

Can I heat Trizol?

I wonder if I can heat Trizol reagent for 30 min 65C. The goal is to disrupt protein-RNA complex while inhibiting nucleases. (I can't use RNasin cause it's inactivated in 65C, and can't use RVC cause ...
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4answers
102 views

Theoretically, what technique would one use to modify a virus so that it only affected a subset of the population?

I'm writing a novel and i would like to know some of the equipment and techniques involved with modifying a virus. Is it feasible for a virus to be engineered to only affect certain people? It ...
5
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1answer
69 views

No digested product bands, but markers are visible. What could be the reasons?

I had to digest my insert containing plasmid and another vector into which my insert has to be ligated, each with BamHI and NotI. Upon digestion, I ran the gel to check for results. The digestion ...
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1answer
84 views

The mechanism of mechanoreception?

I am interested in knowing the molecular mechanism behind mechanoreception/mechanotransduction (i.e. mechanism behind receptor potential generation on mechanical stimulation). I know that most ...
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3answers
74 views

Hebbian theory “fire together” clarification

Donald Hebb states it as follows: "Let us assume that the persistence or repetition of a reverberatory activity (or "trace") tends to induce lasting cellular changes that add to its stability.… ...
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0answers
30 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 ...
5
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2answers
51 views

What are some atypical examples of positive transcriptional cooperativity?

Cooperativity in gene expression is an important feature of many regulatory networks. Described using the Hill function, the most common example is a transcription factor (TF) that when bound to its ...
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59 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
5
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1answer
130 views

High frequency human genetic oscillators?

The most well studied genetic oscillators in human genomes are involved in regulating the circadian clock (which operates on an approximately 24-hour cycle) and cell cycle activity (with single cycles ...
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1answer
137 views

RNeasy Mini Kit low 260/230 ratio — can I purify this RNA for further use?

I used Qiagen's RNeasy Mini Kit to isolate RNA from 5*10^5 C28/I2 (immortalized human chondrocytes). However, my yield is low (~25 ng/ul), but my 260/280 ratio is great (~2.3), and my 260/230 ratio is ...
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19 views

coevolution by physical interaction between RNA sequences

There are several items that relate to the concerted evolution between pairs of sencuencias (especially structural) rRNA as a consequence of physical interaction in tertiary, or even quaternary ...
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19 views

detection of primary origin (natural selection or genetic drift) on alterations in sequences

I recently had a question about how to distinguish the origin of variations in the sequences of rRNA as to whether alterations are generated by genetic drift or natural selection. For me it has not ...
2
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1answer
623 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 ...
3
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1answer
111 views

Optimization of annealing temperature

I am trying to optimize annealing temperature for some primer pairs. I have tried optimization using cDNA, genomic DNA, Taq polymerase, phusion polymerase etc., but every time I am either getting ...
3
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1answer
45 views

Efficiency of plasmid DNA isolation from frozen E. coli cell cultures

Has anyone isolated plasmid DNA from frozen (at -20degrees) E. coli cell cultures (not pellets)? Has that worked and if so, with what yields? What would be the quality of the isolated plasmid DNA if ...