Messenger RNA is produced during transcription before it leaves the nucleus to be be translated by a ribosome. This produces a sequence of nucleic acids for which mRNA is considered the template.

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Multiple Genes per mRNA [closed]

I know that in prokaryotes, there are regularly multiple genes on a single strand of mRNA. I couldn't find any information on what would happen in a eukaryote, however. On one hand, the stop codon ...
4
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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. ...
8
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2answers
104 views

How to find miRNA binding sites on a specific gene?

I am trying to find miRNAs that bind to the 3'UTR of a specific gene. What is the best way of doing that (that is, with a good scoring analysis that is most commonly used by researchers in this area)? ...
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33 views

Artificial Gene Alteration [closed]

Is artificial alteration/"correction" of genes conceivable? I know viruses can manipulate DNA or RNA to replicate itself, but its only temporary, right? If this were possible, how would it be gone ...
4
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1answer
185 views

How many RNA-binding proteins can simultaneously bind on a single mRNA?

Typically, how many RNA-binding proteins can simultaneously bind to a single mRNA? Or said differently, how many "binding sites" does an mRNA have? What order of magnitude? I am interested in RNA ...
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10 views

How to automate this custom gene expression microarray chip?

I have 16 gene sequences that I am interested in their detection in clinical samples. I already measured their RNA levels by using RT-qPCR technique and their benefit has already been proven to help ...
3
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1answer
73 views

Remove RNAse from pipettes?

Our lab uses old Gilson Pipetman pipettes. I've been using some for making mRNA by in vitro transcription and started running into RNAse issues. I'm trying to clean everything up, including the ...
2
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2answers
35 views

Valid comparison of gene expression between several genes in several cell lines

We have gene expression data (Affimetrix mRNA gene expression results) for several cell lines, over a set of genes. Our goal would be to be able to compare relative gene expression for genes over the ...
3
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50 views

Circular mRNA to produce long proteins

Ribosomes can read mRNA and produce proteins, if we somehow make a circular mRNA for the ribosome to bind onto, it will make infinitely long "proteins", (since ribosomes can make very big proteins, I ...
4
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1answer
70 views

Ribosomal RNA amount in a Drosophila cell

I am isolation RNA from Drosophila larvae brain with TRIzol method. What percentage of extracted RNA will be ribosomal RNA? I am only interested in mRNA, so I am trying to figure out whether I need to ...
3
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1answer
56 views

mRNA with Pseudouridine and 5-methylcytidine?

I am making mRNA by in vitro transcription for transfection purposes. There is a lot of literature about how protein expression is increased and innate immune activation is decreased by substituting ...
8
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2answers
2k views

why is AUG the initiation codon?

Is there any reason why AUG is the initiation codon ? Why is there a need for an initiation codon ? Can't translation start with different codons?
2
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1answer
56 views

Mutation in pre-mRNA sequence

Has there any mutations been recorded which cause harmful effects due to change in the part of pre-mRNA responsible for proper m-RNA splicing ?
2
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3answers
386 views

How is the mRNA transported out of the endoplasmic reticulum?

In eukaryotes the nuclear envelope is continuous with the ER, so what helps it out of that?
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77 views

Is the poly(A) tail added while transcription is still underway?

I read in Campbell that ....splicing and poly A tail addition may also occur while transcription is still under way. How can the poly(A) tail be added while transcription is still going on?
4
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2answers
181 views

Current state of direct RNA sequencing

I had a colleague ask me recently whether mRNAs could be sequenced directly. I found this Nature paper[1] published by Helicos in 2009, in which they describe their developments in the area. It's been ...
2
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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.
4
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1answer
65 views

What is the advantage of the way eukaryotes initiate translation?

The eukaryote and prokaryote mechanism for translation is slightly different. Is there any advantage of the eukaryote translation mechanism ? Edit : I specifically want to know why eukaryotic ...
7
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2answers
455 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 ...
2
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1answer
164 views

How can a continuous RNA be transcribed in the lac operon?

The lac operon has 3 genes: lacZ , lacY and lacA. I have seen that the m-RNA transcript of these genes has stop codons in between. So, how can the RNA be made continuously? Won't the RNA Pol detach if ...
5
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2answers
263 views

How to find Exons in mRNA Computationally

I'm having trouble finding a method to find exons in the original DNA sequence used to create the mRNA, even given the sequence of the mRNA, as I cannot find a way to reliably identify the beginning ...
7
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1answer
84 views

Do antisense transcripts have different names than their sense strand transcripts?

I want to find which genes in the human genome can potentially be complementary to a transcript that could act as antisense transcript inhibtion? Are cis-NATs (naturally occuring anti-sense ...
18
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2answers
3k views

Do gene expression levels necessarily correspond to levels of protein activation?

I have seen a lot of research into molecular mechanisms of diseases/phenotypes use measures of RNA as a 'proxy' for the level of protein available in the cell. Is this actually valid? My problem ...
15
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3answers
408 views

Is exon order always preserved in splicing?

Are there any cases in which the splicing machinery constructs an mRNA in which the exons are not in the 5' -> 3' genomic order? I'm interested any such cases, whether they involve constitutive or ...
7
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1answer
106 views

How can I normalize mRNA samples for sequencing?

Is there an easy, inexpensive, not too labor intensive way to normalise mRNA samples so that even though one loses information of gene expression levels, each of the transcripts in the transcriptome ...
2
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97 views

Has anybody used Evrogen's DSN-normalization protocol for cDNA normalization? [closed]

I found the link to a commercial product by Evrogen to normalise cDNA samples for gene discovery projects here: http://www.evrogen.com/technologies/normalization.shtml The most up-to-date reference ...
9
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1answer
91 views

Which patterns do I have to avoid when modifying the 3'-UTR?

I want to change a pre-miRNA sequence (in my case the pre-miRNA is encoding in a 3'UTR of a gene) and then put it in a lentivirus to see if it is still processed. After modification (permutation of ...