how to perform rna sequencing?
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['Design Experiment. Set up the experiment to address your questions.', 'RNA Preparation. Isolate and purify input RNA.', 'Prepare Libraries. Convert the RNA to cDNA; add sequencing adapters.', 'Sequence. Sequence cDNAs using a sequencing platform.', 'Analysis.']
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['Find MFOLD on the web. Enter this URL for MFOLD: ... ', 'Then click the RNA Folding Form link.', 'Which will open up the following form: ... ', 'Optional Step. ... ', 'After a short wait, you will get this output page. ... ', 'ENERGY DOT PLOT. ... ', 'The dots in the box correspond to the indicated helical stem region in the structure. ... ', 'ENERGY DOT PLOT.']
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Long non-coding RNAs (lncRNAs) are grouped into transcripts that are > 200 nucleotides in length. The human genome is estimated to contain ~16,000 lncRNA genes (Gencode 27). ... Most of the lncRNAs contain normal 5'-caps and 3' poly-A tails.
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['Extracting the DNA from cells.', 'Cutting up the DNA using an enzyme.', 'Separating the DNA fragments on a gel.', 'Transferring the DNA onto paper.', 'Adding the radioactive probe.', 'Setting up the X-ray film.', "Yes - we've got the result!"]
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Transcription by RNA polymerase. An RNA strand is synthesized in the 5′ → 3′ direction from a locally single stranded region of DNA.
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All of the RNA in a cell is made by DNA transcription, a process that has certain similarities to the process of DNA replication discussed in Chapter 5. Transcription begins with the opening and unwinding of a small portion of the DNA double helix to expose the bases on each DNA strand.
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Ribosomal RNA is transcribed from ribosomal DNA (rDNA) and then bound to ribosomal proteins to form small and large ribosome subunits. rRNA is the physical and mechanical actor of the ribosome that forces transfer RNA (tRNA) and messenger RNA (mRNA) to process and translate the latter into proteins.
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Three major types of RNA are mRNA, or messenger RNA, that serve as temporary copies of the information found in DNA; rRNA, or ribosomal RNA, that serve as structural components of protein-making structures known as ribosomes; and finally, tRNA, or transfer RNA, that ferry amino acids to the ribosome to be assembled ...
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RNA is then translated into proteins by structures called ribosomes. There are three types of RNA involved in the translation process: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). Although some RNA molecules are passive copies of DNA, many play crucial, active roles in the cell.
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Most splicing occurs between exons on a single RNA transcript, but occasionally trans-splicing occurs, in which exons on different pre-mRNAs are ligated together. The splicing process occurs in cellular machines called spliceosomes, in which the snRNPs are found along with additional proteins.
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The first step in shotgun sequencing an entire genome is to digest the genome into a large number of small fragments suitable for sequencing. All the small fragments are then cloned and sequenced. Computers analyze the sequence data for overlapping regions and assemble the sequences into several large contigs.
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RNA is synthesized from DNA by an enzyme known as RNA polymerase during a process called transcription. The new RNA sequences are complementary to their DNA template, rather than being identical copies of the template. RNA is then translated into proteins by structures called ribosomes.
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['From existing DNA to make new DNA (DNA replication?)', 'From DNA to make new RNA (transcription)', 'From RNA to make new proteins (translation).']
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['Polymerase Chain Reaction (PCR) ... ', 'Restriction Enzymes (Molecular Scissor) ... ', 'Gel Electrophoresis. ... ', 'DNA Ligase. ... ', 'Plasmids. ... ', 'Transformation/Transduction. ... ', 'Identifying Transgenic Organisms.']
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Transcription results in production of RNA , it can be mRNA , rRNA and tRNA.
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Transcription is the process by which DNA is copied (transcribed) to mRNA, which carries the information needed for protein synthesis. Transcription takes place in two broad steps. First, pre-messenger RNA is formed, with the involvement of RNA polymerase enzymes.
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Of the many types of RNA, the three most well-known and most commonly studied are messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA), which are present in all organisms. These and other types of RNAs primarily carry out biochemical reactions, similar to enzymes.