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1

Reliability comes from the fact that in Sanger sequencing (AFAIK) the synthesis of the new strand to the template and the actual reading of the sequence is separated. As you may know Sanger seq uses capillary gel elfo. thus separating the newly produced DNA fragments on a single nucleotide difference, and the reading is done by fluorescence detectors (then ...


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Being no expert on pain, I will share some thoughts on the issue. According to the following site(http://www.helpforpain.com/arch2000dec.htm), there are two types of pain: nociceptive and neuropathic pain. Neuropathic pain involves the central and peripheral nervous system, a possibility I would discount due to no apparent link to suffocation. Thus, it is ...


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If you have the money to buy a kit, the QIAmp kit has always performed well for me.


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If by enzyme you mean "protein" aka polypeptide, than there are such things as catalitic RNAs. Those are molecules of RNA that facilitate chemical reactions but don't change themselves (definition of catalyst). I think that, based on the discovery of such RNAs, it is now believed that life might have started from or with the help of catalytic RNAs (please ...


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Given the fact that the negative control uncut plasmids are producing white colonies, it seems that there have been significant mutations within the lacZ gene in the plasmid, causing the uncut plasmids to become white due to inactivation of the gene. I would suggest purchasing a new batch of pBluescript, or if that is not possible for whatever reason, ...


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It depends both on structure and charge. Binding sites of proteins are essentially formed by amino acids placed in a particular conformations such that it will match the binding site of their counterpart protein or substrate. This is commonly referred to as the lock and key model of protein binding. This is similar to how two puzzle pieces fit together, the ...


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Short answer is that higher temperature favors annealing of longer sequences. There are number of ways to calculate melting temperature, but all of them produce similar results: longer polymers require more thermal energy to melt. Hence, quick cooling from higher (say, from 95C thermocycler can cool in 10-12 sec) to RT/4C will favor re-annealing of circular ...


5

I've found an old paper using radioactively labeled bacteria to track their fate in macrophages. According to the data, both $^{14}$C and $^{32}$P are reutilized in the host cell. COHN ZA. (1963). The fate of bacteria within phagocytic cells. I. The degradation of isotopically labeled bacteria by polymorphonuclear leucocytes and macrophages. J Exp Med. Jan ...


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In this article it is described that starved macrophages 'consume' heat inactivated bacteria through phagocytosis with enhanced ability. Starving can also induce (macro)autophagy and the pathways are connected with phagocytosis,in the macrophages under investigation it was shown that autophagy did not play role in the enhanced ability of phagocytosis. The ...


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Deinococcus radiodurans did not "develop resistance to mutations". It is able to repair its chromosome when scatered in pieces by radiations or desiccation, while other bacteria would die in such conditions. So this is adaptive in extreme environments, such as deserts (where it has evolved) or canned corned beef (where it was discovered).


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Thermo Fisher has a protocol for the separation of host mammalian RNA from prokaryotic RNA, optimized for E. coli vs human, mouse or rat sequences. Capture oligonucleotides bind portions of the mammalian RNA and hybridize. These hybridized oligo/RNA are then removed from solution via "oligonucleotide-derivatized magnetic beads," after which the remaining RNA ...


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In my experience, transduction efficiencies skyrocketed when I applied a spin-infection procedure. A 30 min low speed centrifuge with the virus changed a lot. Here is the TRC protocol for this: http://www.broadinstitute.org/rnai/public/dir/download?dirpath=protocols/production&filename=TRC%20viral%20infection%20200909.pdf


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The question is following: did you incubate cells on ice before spinning? Protocol, for example this one for $DH5\alpha$, calls for just 10 minute incubation after heat shock: Heat shock at exactly 42°C for exactly 30 seconds. Do not mix. Place on ice for 5 minutes. Do not mix. Pipette 950 µl of room temperature SOC into the mixture. Did you ...


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This is hard to answer since we don't know the exact conditions you chose, as these are rotation speed (or better g), how long you spun them down and also if you used a cooled centrifuge or not. It's also important how gentle you resuspended the pellet. However, I would plate these cells, no matter what happened since you are at the last step of the ...


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I have looked about the topic and I have asked experts and the best aproximation to know the vertebrate host from arthropod blood meals is targeting mitochondrial genes because you can get clean sequence data due the fact that only one allele is present rather than two. Within the mitochondrial genome, you should select a gene which has been widely ...


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So, as I pointed out in comments, and requoting: "The direction in which cargo is transported is dependent in part upon the position of the motor domain, which can be located N-terminally (N-kinesins), C-terminally (C-kinesins) or internally (M-kinesins). In general, kinesins with N-terminal motor domains move their cargo towards the plus ends of ...


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I wanted to illustrate the process I explained in a comment on aandreev's answer with some images, I just copy/pasted the actual text sections, however. From comments: The tRNA is bound to an elongation EF-Tu (which is also bound to GTP). The Ef-Tu has an affinity for a factor binding site in the A-site of the large ribosomal subunit. You have an anticodon ...


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As with many processes in cellular biology, nothing really moves anywhere, attracted by specific force. What that means, is that any molecule has a chance to interact with any other molecule, say, bind to it. Only force that is acted is electrical force of attraction of negative charges to positive ones. Due to diffusion and temperature fluctuations, ...


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No. DNA, and RNA have a polarity in the orientation of their sugar-phosphate backbones, based on the numbering of the carbon atoms in the sugar rings. So, for example, we say that the sequence of this RNA is being read from the 5' to the 3' end, which indicates that the first nucleotide is exposing the hydroxyl at the 5’ carbon on its sugar ring. At the ...


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Cytoskeletal nucleation refers to the de novo formation of cytoskeleton filaments. If microtubules are nucleating, that means microtubules are being polymerized from lit. "From the beginning" by α/ß-tubulin dimers. So literally, and we see in the image below, Arp2/3 complexes act as progenitor sites so to say for new actin filaments by the "nucleation" of ...


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In this context, i feel nucleated means being a "core structure", a substrate for the organization of one or multiple polymer chains.


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The answer is in the slides you provided a link to. The key fact is that Craig & Andy did careful controls to show that it was the presence of dsRNA in both the sense control, and in the anti-sense experimental sample that was responsible for the RNAi-mediated knock-down. Actually, Ken Kemphues showed this earlier in a Nature paper (the control also ...


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Mitochondria are granular or filamentous organelle, which are present in the cytoplasm of a cell. They are ovoid in shape, with the presence of double membranes. Mitochondrial fusion is required to distribute mitochondrial DNA to the mitochondrial population and to maintain its respiratory competent and energized organelles. Mitochondrial division is ...


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Memories are not stored in just one part of the brain. They are widely distributed throughout the cortex and neurons. The long-term memories are stored throughout the brain as groups of neurons, the brain stores memories in three ways--Short-term memories, Sensory memory and Long-term memory the human brain cell can hold 5 times as much of information as ...


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I assume it has an analogous function in ligation buffers. There it apparently takes up a large proportion of the volume and thereby increases the chance of interaction between bits of DNA. In a transformation buffer it should increase the chance of DNA getting into a cell. Unfortunately the only reference I found for this was at Bitesizebio: ...



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