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

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2k views

Very high 260/230 absorbance ratio of an RNA sample

After my most recent RNA extraction, the RNA samples had very high 260/230 absorbance ratios, (ranging from 5 to 25). I've never gotten numbers like this and I know the ratio is supposed to be ~2 in ...
11
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3answers
3k views

How long can E. coli stocks be stored at -20°C?

I'm volunteering for a biohacker lab - biocurious in Sunnyvale. The have a pretty good set of equipment - gel boxes, incubators, but they don't have a -80°C freezer yet. I'd like to set up some ...
1
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1answer
37 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 ...
2
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1answer
89 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 ?
2
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1answer
215 views

How can Southern blotting be used to establish gene copy number?

I am reading a paper on a certain marine microorganism that can eat alkanes and therefore might be useful for cleaning up oil spills, etc. This sentence struck me as odd (although I am sure that the ...
3
votes
1answer
150 views

What are centromeres *really*?

I've gathered that a centromere is a a region* where the DNA is bundles up even tighter (around protein different to Histone) and chromatids are 'joined'. However I'm still mostly in the dark ...
19
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1answer
486 views

Regulation of chromatin structure

Recently, I reviewed the different levels of chromatin structure. The primary level is nucleosomes, where DNA is bound to histones, and has structural similarity to "beads on a string." The secondary ...
10
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1answer
417 views

What is the functional and structural distinction between core (H2A, H2B, H3,H4) and linker(H1/H5) histones?

Many explanations of histone biochemistry isn't quite elucidating for the undergraduate student. How does histone structure (dimers, octomers) relate to their specific functions as core or linker ...
2
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1answer
130 views

Histone marks mechanism

I am slightly confused about the mechanisms that makes histone modifications associate with gene expression. That is, H3K36me3 is believed to be present in actively transcribed genes, H3K27me3 in ...
9
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4answers
3k views

Why we have no enzyme to digest cellulose?

As we know, cellulose is the most abundant polysaccharide in nature. Why don't we have any enzyme to digest cellulose?
0
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1answer
49 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 ...
3
votes
1answer
79 views

Transposons and the net amount of DNA

The retrotransposons and certain DNA-transposons, are "jumping" sequences which may be incorporated elsewhere in the genomic DNA of an organism, through varying mechanisms. This insertion is almost ...
1
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1answer
265 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 ...
2
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2answers
68 views

Molecular simulations without capping n- and c-terminus of polypeptide

When running molecular simulations, what is the consequence of not capping the n- and c-terminus? I have been following gromacs tutorials and haven't come across any that deal with capping the ...
8
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2answers
192 views

How are antibodies designed?

Antibodies have the ability of recognising highly specific peptide sequences and bind it at their antigen-binding site. This ability is harnessed as a tool in research to purify target structures in ...
2
votes
1answer
74 views

Determining sequence of oligoribonucleotide

Oligoribonucleotide X was treated with phosphatase (for removal of 3' and 5' - terminal phosphates), then with RNAase T1, which cleaves all phosphodiester bonds located in a 3' position of ...
1
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1answer
55 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 ...
3
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2answers
78 views

Why not use Cu(OH)2 as a dye for protein electrophoresis?

As I know, $Cu(OH)_2$ $+$ Protein $\rightarrow$ Purple. So why not use $Cu(OH)_2$ as dye in protein electrophoresis?
4
votes
1answer
271 views

How much does the Hayflick limit/telomere length vary across taxa and within humans?

Since they seem to be quite connected, I'm curious if anyone knows of research comparing the Hayflick limit (and presumably by extension telomere length) between different taxa. I've heard the ...
1
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2answers
73 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 ...
1
vote
1answer
59 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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0answers
1k 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 ...
0
votes
0answers
29 views

Where can I get yeast mutants? [duplicate]

I want to use some yeast mutants to conduct gene functional analysis, and the gene was isolated from plants. Can anybody give me some information regarding the websites or agencies from which I can ...
0
votes
1answer
80 views

Hypothetically, what is the smallest functional form of human?

Since the my first encounter with doraemon manga, I've been wondering many things, one of them is the small light. Assuming we had an instrument, such as small light, which enabled people to grow and ...
1
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2answers
348 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 ...
2
votes
1answer
86 views

What will happen if we expose the brain to an intermittent light?

If a brain is exposed to an intermittent light are specific areas going to fire? If yes, which of them? Is there any experiment about this?
1
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2answers
101 views

Protein purification methods

Which of the following methods would yield the most purified protein fraction? A. Salt precipitation B. Charge separation C. Affinity purification The first one is the most commonly used, is it ...
0
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1answer
63 views

Electrophoretic mobility of isozymes

If isozymes are seperated using electrophoresis, which of the following will be the principle of separation ? A.charge density B.molecular weight C.polarity of molecule I think it should be B.
0
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1answer
46 views

Proportion of amino acids from a random polymer of U and C

A protein synthesis assay was carried out in vitro. A polyribonucleotide containing U and C in proportion 1:5 (positions of U and C are random) was used as a template. Which amino acids and in ...
3
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3answers
177 views

How to learn molecular biology through pubmed research articles?

Instead of using a textbook, is there an alternative curriculum, that simply lists a set of pubmed research articles for each topic covered in a typical undergrad molecular biology course? I am ...
1
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1answer
146 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.
2
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0answers
43 views

Shortest route for a Nitrogen atom from fossil fuel to a protein in your body? [closed]

Millions of years ago fossil fuels originated from plants. A nitrogen atom from fossil fuel is now inside in a protein molecule in your body. What is the shortest path that the nitrogen atom ...
5
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3answers
179 views

What does it mean for a distribution to be “consistent with a two rate-limiting stochastic steps”?

I'm reading a study (full text here) that examine the dynamic of nuclear translocation of a transcription factor in budding yeast, in response of calcium stress. They found that it occurs in bursts, ...
3
votes
1answer
857 views

How does blue white screening work in selecting E coli that have taken up a plasmid vector?

The plasmid vector I am referring to is pCR 2.1 - TOPO. I added the vector to the E coli and plated them up onto LB+amp+X-gal plate, then incubated. After incubation the plates had two types of ...
4
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2answers
395 views

Does bleach destroy RNAse activity, and if so, how does it do it?

I am working with RNA samples, and I'm trying to be very careful about RNAse contamination. I have some questions about bleach, though. Some people say that a solution of bleach is enough to destroy ...
3
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0answers
38 views

Adenosine metabolism

Are adenosine or its catabolites increased in inflamed airways? How can I assess this? I am trying to use inhibitors for adenosine deaminase, xanthine oxidase, and purine nucleoside phosphorylase, but ...
17
votes
3answers
1k views

How long can I store extracted RNA?

If I extract RNA from a (leaf tissue) sample using a one-step phenol:chloroform extraction, how long can those samples be stored at -80°C? And how many times can I defrost and refreeze them before ...
8
votes
2answers
92 views

Is there a Reverse Transcription optimization for long, 9kb, transcripts?

Has anyone optimized RT for long transcripts (9kb)? The downstream application will be PCR amplification and Illumina library prep. It will be trivial to make internal primers sets for the PCR that ...
6
votes
4answers
565 views

Does RNA polymerase move around DNA or does DNA rotate benath the polymerase?

I'm thinking of the human genome specfically, but more general answers are welcome. As RNA polymerase moves along the DNA helix it follows a single strand. The two DNA strands are unwound locally ...
4
votes
1answer
173 views

Cloning two fluorescent proteins with different promoters in the same plasmid?

I want to clone two fluorescent proteins both driven by different promoters in a plasmid to be used to transform B. subtilis at the amyE locus. For this I want to use plasmid pSG1729 (Lewis and ...
1
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1answer
77 views

Do mitochondrial DNA have repair enzymes?

Mitochondria are considered endosymbionts originated from a prokaryote. So, do they have a DNA repair mechanism ?
3
votes
1answer
490 views

Why re-label the strand ends in 3' DNA labelling?

I have a problem with a molecular biology question; I don’t understand how DNA 3’ labelling works. I took a diagram from my lesson and tried to understand with it; this is what I understood. If I’m ...
5
votes
2answers
108 views

There are linear and rotary molecular motors in the cells. Do any of them have a fixed or stable frequency or speed?

Are there any linear, rotary or oscillatory molecular motors in the cells which can have fixed frequeny and which can be used as a reference for elapsed time timer? This question is with relevence to ...
2
votes
2answers
72 views

Termination of translation

What dissociates first - the last tRNA, mRNA and release factors or the subunits of ribosomes? I tried searching this from Lehninger but couldn't get a clean answer.
5
votes
1answer
138 views

Synthetic construct with multiple ORFs not expressing

I designed a synthetic construct on paper and got it synthesized from a company. The objective was to make a vector which can be used to study both transcriptional and post-transcriptional regulation ...
3
votes
1answer
1k views

What is the purpose of Y-shaped adapters in Illumina sequencing?

Y adapters different sequences to be annealed to the 5' and 3' ends of each molecule in a library. The arms of the Y are unique, and the middle part, connected to the DNA fragment, is complementary. ...
7
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2answers
490 views

What exactly happens if during translation, an amino acid is not present?

Lets say that the cell wants to make a particular protein. Transcription of the appropriate gene is done and the mRNA is made. mRNA attaches to the ribosome and the translation is initiated in a ...
1
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2answers
43 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 ...
1
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1answer
921 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). ...
1
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1answer
120 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 ...