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

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3
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0answers
22 views

What are flagged primers? [closed]

I'm interested in amplifying a sequence for further use with Gibson Assembly. I want to create overhang regions in my DNA fragment so there would be complementarity to the plasmid I'm trying to insert ...
1
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0answers
19 views

Yew seed dormancy reduction

Yew seeds have very long dormancy when artificially planted. They are also adapted to passage through digestive systems of animals that eat/transport them. But could the dormancy be shorter (or the ...
3
votes
3answers
78 views

miRNA targets mRNAs

If we observe that a miRNA, say hsa-mir-233, targets a mRNA, say XXX, in a given tissue in disease condition, can we say that always hsa-mir-233 targets mRNA XXX regardless of type of tissue and ...
1
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0answers
82 views

Why do PCR manufacturers recommend assembling the reaction mix on ice?

Many PCR manufacturers recommend preparing the reaction mix for the reaction on ice. For example, here is NEB's recommendation for their Q5 polymerase. Other manufacturers also include similar steps: ...
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1answer
37 views

gene transformation from plant to plant - transgenic plant

I want to know if the expression of a transfered gene depends on source of the gene, if e.g.: 1] I isolate a gene from plant and transfered it to the same plant? 2] I isolate a gene that is highly ...
2
votes
1answer
25 views

Degree of complementarity

Apologies in advance for asking bit naive question. I have looked up for this concept a lot but didn't find anything. The wikipedia (I know its not authentic all the time, but I think works well for ...
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1answer
97 views

Enzymes and plasmids

A high school class has analyzed plasmids by the help of restriction enzymes and electrophoresis. The class got delivered two different plasmids, pBR 322 and pC 508. These two plasmids were going to ...
-2
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1answer
76 views

Homework Question: Confused about figure related to cDNA and gDNA

what is the answer in blank? I think the loop structure for the top blank is an intron. I think RNA is the answer for the bottom blank, but it is cDNA, so it can't be RNA. I cannot figure out what ...
2
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1answer
122 views

What does it mean to clone a gene?

When I look up information related to the identification of disease genes, texts will often refer to the gene being "first cloned." What does "clone" mean in this context? Is it simply a synonym for ...
2
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0answers
50 views

PCR that worked previously is now only showing primer dimers and a smear on gel

PCR amplification of a promoter sequence for gel extraction worked beautifully using Phusion HF enzyme with GC (higher error but less picky) buffer. However, DNA concentration from the gel extraction ...
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0answers
18 views

Amputation of part of the cytoplasm of the amoeba? [closed]

We ampute a part of the cytoplasm of the amoeba many times and we observe that amoeba is not divided This experience allows to deduce the cause of the division of amoeba Do that have relation with ...
2
votes
1answer
39 views

genetic complementation problem

I've attached the picture of the problem. SO, based on the information I managed to assemble two large cistrons. (if the combination of two mutants yield (-) mutations are on the same chromosome) A)...
1
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0answers
38 views

Can genes be expressed sequentially?

As I understand it, any gene on an exposed/unpacked region of a chromosome is continuously being expressed. Regulatory genes may increase or decrease the amount of protein synthesised due to its ...
1
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0answers
34 views

Biological modeling languages - some basic issues

I am trying to get used to with couple of modeling languages in biological context like SBML, BioPAX, Kappa and BEL. Some basic issues came up to my mind that are unclear to me even after a lot of ...
2
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0answers
33 views

Gel Electrophoresis

we add Etbr within the agarose gel but we do not add Etbr directly in native PAGE ?when we make agarose gel, we add Etbr before its solidification but we do not add Etbr when the native PAGE gel is in ...
1
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3answers
44 views

importance of cDNA sequencing of a specific gene in different plants

i want to know what if i made a sequence of cDNA of a specific gene in different plants - what is the important from the sequence and can if found difference between plants , and can i use this ...
2
votes
2answers
114 views

Why does agrose gel runs horizontally, and SDS-PAGE gel runs vertically?

It seems in electrophoresis, agrose gel always runs horizontally, and SDS-PAGE gel always runs vertically, why?
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0answers
12 views

Function of pseudogenes and its process through translation

Can pseudogenes be translated and function properly or will it be seen as a mutation instead?
2
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2answers
73 views

How important is non-binding site of protein?

Just a curious question: What will happen if i remove most protein sequence that encoded for non-binding site of Restriction Enzyme to make a new enzyme, can the new restriction enzyme still work?
2
votes
1answer
2k views

What is the role of glycine in the running buffer for SDS-polyacrylamide gel electrophoresis

We used pH6.8 in stacking and pH8.8 in resolving gel. In the class, the professor explained that the glycine change is like: ...
3
votes
2answers
124 views

Question about proto-oncogenes and oncogenes?

My textbook says: Growth-promoting genes are called proto-oncogenes. Some can be changed into oncogenes by a point mutation that alters the ability of the proto-oncogene to be switched off. They ...
5
votes
1answer
141 views

How is adrenaline a ligand?

I keep reading that adrenaline is a ligand, however from what I understand a ligand is a molecule or ion which donates a pair of electrons to a central transition metal ion in a complex. How then is ...
1
vote
1answer
76 views

Is there any way to make protein pass through cell membrane?

Protein cannot pass cell membrane because it's a large molecule. Until now, is there any technique that can make protein pass through the cell membrane in vivo? I want to create a protein-drug that ...
1
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2answers
91 views

Getting PCR amplification at annealing higher than Tm!

I am amplifying a gene where in a gradient pcr i am getting amplification at an annealing temperature about 5 degrees (67) higher than Tm (62.5)? What is wrong here? Also, I am getting a very strong ...
0
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2answers
45 views

General transcription factors vs. DNA Binding transcription factors

I know that DNA binding transcription factors are trans-acting, but what about general transcription factors? Are they cis or trans acting?
0
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0answers
40 views

What is the difference between basal transcription and activated transcription?

My professor keeps mentioning these two forms of transcription and I'm completely confused as to what the difference is between them. I didn't even realise there were two different types of ...
1
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1answer
47 views

Can we create custom gene/protein?

Does it possible to create any custom gene or protein we want with current technology? I have a protein sequence or a gene sequence about 4000 bp write down on my computer, is there anyway to "print" ...
1
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0answers
23 views

What is the difference between cell, membrane and animal based assay?

I am reading a review about MRP1, discussing it's ability to extrude different compounds from cell interior. When describing the different experiments conducted to observe the MRP1's reaction to ...
3
votes
1answer
136 views

In CRISPR bacteria, how does viral genomes get integrated into the spacers of CRISPR? Also, in its use, where does Cas9 cut the DNA?

I've been out of Biology for about a year polishing my programming skills. I know CRISPR/Cas9 allows targeted 'cutting' of DNA via RNA-guidance. Few questions regarding this. Regarding to its ...
0
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2answers
199 views

Electroporation vs gene guns [closed]

What are the pros and cons of using electroporators (left) and gene guns (right) for transformation in terms of: Price Target organism Efficacy Ease of use Maintenance
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0answers
37 views

Is pathlength correction vital to be included while quantifying protein

I am trying to quantify protein in serum/plasma samples. I have carried out biuret assay for my samples using microplate reader. While performing I encountered the doubt whether I should include ...
2
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0answers
40 views

Gene sequence and DNA [closed]

can anyone help me with this? It shows two gene sequences for a sheep and a goat. ...
2
votes
1answer
108 views

What is the molecular mechanism of odorant transduction signaling pathway?

I have read a few papers on the topic of odorant signaling pathway. The following papers are listed as examples. However, I found that there is not a clear pathway representation of the odorant ...
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0answers
34 views

Wash adsorbed protein without destroying Biotin-Avidin-Mechanism

I am looking for reliable protocols to wash protein (e.g. Fibronectin) adsorbed onto glass surfaces. According to Protein immobilization literature, people usually use 1% SDS and some incubation time. ...
3
votes
1answer
91 views

What is the shortest mRNA the ribosome can read to produce a peptide?

This question came as a comment on a previous question regarding non-ribosomal peptide synthesis, and why Glutathione cannot be synthesized by the ribosome. In general, Glutathione has a "gamma" ...
1
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1answer
33 views

Conjugated deoxyribonucleotides

I'm currently learning about using PCR techniques to make fluorescently labelled DNA probes, and the textbook mentions "conjugated deoxyribonucleotides" Can someone explain what these are? Nothing ...
2
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2answers
549 views

Why is beta actin commonly used as a control in Western blots?

It is about the western blot question. The paper:http://www.sciencedirect.com/science/article/pii/S1931312813001145. In the western blot experiment in this paper, they use beta actin as control. And ...
5
votes
2answers
143 views

Non-ribosomal peptide synthesis: why Glutathione cannot be produced by the ribosome?

Case: I am writing a summary for a class in protein structure and function, and was asked to describe some different ways that peptides are synthesized (that does not involve the ribosome). I ...
1
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1answer
179 views

What is the reason for having an extra recognition site for a restriction enzyme?

Can the size of a supercoiled plasmid DNA be determined by using standard DNA size fragment electrophoresed in parallel? 2. An unknown DNA molecule was cleaved using several restriction enzymes ...
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0answers
23 views

If two different diseases have more than one common pathway, the relationship between their occurences?

I just wonder that are there any special relationship between the pathway and disease occurrence. For example, there are cancer A and cancer B (and they have more than one common pathway). Then if ...
4
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1answer
54 views

Function of NaCl in yeast shuttle prep of plasmids

What is the role of NaCl in plasmid isolation from yeast cells using plasmid rescue solution? If the basic theory behind the method is alkaline lysis, then we is NaCl used and not NaOH?
2
votes
1answer
115 views

How does one insert cas9 into animal cells?

How could cas9 be inserted into cells by researchers looking to edit a genome? I imagine for engineering bacterial systems you could just put in the cas9 coding region in an expression vector, but is ...
3
votes
1answer
60 views

How to predict a mRNA secondary structure with a large sequence?

When I use some web servers to predict a mRNA secondary structure, I find they always required in a small size sequence. If I use a long sequence and cut it into small parts, do these small parts ...
0
votes
1answer
65 views

On which amino-acids residue is the SDS acting on?

I would like to know exactly what is the mechanisme of the SDS, and I would like to know on which amino-acids residue the SDS is acting on. Can you help me please ? Thank you in advance !
2
votes
1answer
58 views

Difference between the P4 and P5 subtypes of P-type ATPases in plants

In plants there are various kinds of P-type ATPases. What is difference between P4-ATPase and P5-ATPase?
1
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1answer
56 views

What is the attacking mechanism of RF on IgG?

Rheumatoid Factor (RF) attacks Fc portion of Immunoglobulin G (IgG), I want to know the underlying mechanism at molecular level. Also, what type of bond or attachment is made by RF and Fc portion of ...
1
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0answers
26 views

How do molecular recognition features work?

The occurrence of relatively short (10–70 residues), loosely structured protein regions bind within longer, largely disordered sequences that were characterized as bound to larger proteins. These ...
1
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1answer
34 views

Producing a genetically modified animal with cell walls [closed]

I'm curious if there has been any genetic experiments transferring cell wall producing genes into the genome of a animal model organism such as the fruit fly (Drosophila melanogaster) or a larger ...
1
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0answers
39 views

Is there a better lysis buffer than this for fungal DNA extraction?

I've being using this lysis buffer for fungal DNA extraction,lysis buffer - 400mM TRis-HCl(pH8.0), 60mM EDTA(pH8.0), 150mM NaCl, 1% SDS and containing 40microgram/ml RnaseA. It is keep giving very ...
1
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0answers
47 views

How are CheY, CheA, CheW, CheZ pronounced in speech?

This is a quick followup question to this question about proteins that play important roles in chemotaxis: How does one pronounce the protein names "CheY", "CheW", etc., in English? My guess would ...