how can you improve the quality of your RNA preparation?
372436
Rules of extraction of  RNA in the laboratory:\n\n1. Keep away any source of cells - dead or alive (well other than the tissue or cells from which you want to prepare RNA)! So keep your bench clean, use gloved hands etc. Use sterile solutions. If possible, use 'baked' containers.\n\n2. Use RNAse degrading reagents (e.g., RNAse away and/or DEPC -see Sigma catalogue; ambion is a company specialized in RNA-related products, you can try their catalogue/website too) in your solutions.\n\n3. Use RNAse free disposable labwares (like tips and tubes) as much as possible.\n\nABOVE steps should take care of the notorius RNAse enzyme that is present in most of the cells and hard to be inactivated.\n\n4. Use 'RNAse-free DNAse' wherever possible. This avoids DNA contamination.\n\n5. Extend your precautions to the gel electrophoretic analysis and storage of prepared RNA also!\n\nRest: Stick to your protocol
932873
Go to "protein" on wikipedia ( put, protein, in address bar ) then scroll down to "synthesis", for the process. It is too  long a story for my typing skills. The wiki article is not too technical.
670087
Practice your spelling and grammar.
925374
transcription in cells makes RNA copies of DNA.
163081
finding out ways to produce enzymes quickly and efficiently
1140466
Improve your spelling.
340538
Because mRNA comprises only 1-3% of total RNA samples it is not readily detectable even with the most sensitive of methods. Ribosomal RNA, on the other hand, makes up >80% of total RNA samples, with the majority of that comprised by the 28S and 18S rRNA species (in mammalian systems). mRNA quality has historically been assessed by electrophoresis of total RNA followed by staining with ethidium bromideThis method relies on the assumption that rRNA quality and quantity reflect that of the underlying mRNA population. Because mammalian 28S and 18S rRNAs are approximately 5 kb and 2 kb in size, the theoretical 28S:18S ratio is approximately 2.7:1; but a 2:1 ratio has long been considered the benchmark for intact RNA. While crisp 28S and 18S rRNA bands are indicative of intact RNA, it is less clear how these long-lived and abundant molecules actually reflect the quality of the underlying mRNA population, which turns over much more rapidly. \n\nVisual assessment of the 28S:18S rRNA ratio on agarose gels is somewhat subjective because appearance of rRNA bands is affected by electrophoresis conditions, amount of RNA loaded, and saturation of ethidium bromide fluorescence ( An improved analytical tool for total RNA analysis is the Agilent 2100 bioanalyzer, which uses a combination of microfluidics, capillary electrophoresis, and fluorescence to evaluate both RNA concentration and integrity analysis  Another advantage is that it requires very small inputs, allowing the user to assay RNA quality in limiting samples.
144456
Step 1: Good research\nStep 2: Good outline\nStep 3: 5 paragraph form\nStep 4: edit my work
1131043
First learn how to construct a sentence and learn how to punctuate
201658
Start off by letting your thoughts flow -- Jot down ideas, even if they're just words or fragments of sentences, don't think about sentence structure, just write down a plot and add on and elaborate.  Then piece them together into sentences and paragraphs and elaborate more.  Let a few other people read it, hear what they think, and use their input to further increase the quality of your story.  \n\nI hope this helps.  =]
466802
High-throughput DNA Isolation Protocol \n\nCTAB Method \n\n1.      Add 80 ul 2-mercaptoethanol to 40 ml CTAB buffer (see recipe below) right before use (0.2%) and mix. \n\n2.      Preheat CTAB DNA isolation buffer to 65°C in a water bath. \n\n3.      Grind freeze-dried leaf tissue (~100mg) in a Mixer Mill for 3-5 min at 30 time/sec with one 3mm steel bead in a 1.1 ml  tube strip from Qiagen\n\n4.      Carefully pour beads out of tubes and add 300 ul CTAB buffer into each tube with a multi-channel pipette (if a plate is used, you can leave beads in until the end of work)\n\n5.      Cap the tube strips and label each strip, caps and rack, incubate the samples in 65 °C water bath for 60-80 min, use shipping tape to secure the rack and lid to prevent caps from openning up while inverting the rack to mix samples.\n\n6.      Mix samples every 15 min by inverting rack 5-10 times to make sure plant tissue mixes well with the buffer. Watch for sample leaking from caps. \n\n7.      Take samples out of water bath, briefly spin sample box in a centrifuge for 1 min at 1500 rpm (balance box weight before spinning), separate caps from tube and put the cap strips on clean paper and arrange them in order so that they can be reused. \n\n8.      Add 300 Chl/IAA (25:1) into each tube with a multi-channel pipette, cap tube with original cap strips, put a plastic board between tube cap and rack lid to make sure every strip cap gets the same pressure from the lid, cover the box with lid, secure the lid with shipping tape. Mix the samples by inverting the box slowly for 15 min (Chloroform can create pressure and cause leaks of chloroform, so the tubes have to be tightly capped).\n\n9.      Centrifuge at 5,700 rpm  for 10 min. \n\n10.  Transfer the aqueous phase using a wide-bore pipette tip (about 200ul) into a labeled clean plate (650 ul plate), add 150 ul of cold isopropanol (-20 °C), securely cover the plate with a plate cover (or cap strips) and mix gently by inverting the plate several times to precipitate DNA. \n\n11.  Recover the DNA by centrifuging at 5,700 rpm (6100g) for 10 min. Discard the supernatant immediately by suddenly inverting the plate and dry it briefly on several layers of paper towels by briefly touching the plate on paper towels two to three times on different positions of towel to remove excess liquid. This step should be very quick (a few seconds). Then turn the plate back upright and air-dry samples to remove excess liquid for ~10 min.\n\n12.  Add 500 ul of 70% Ethanol directly to the pellet, cover the plate, and invert it gently several times to wash  for 5 min. \n\n13.  Centrifuge at 5,700 rpm for 8 min, and quickly discard wash buffer as described previously (check DNA pellet in each well to make sure DNA stayed in the bottoms of wells\n\n14.  Air-dry DNA pellet at room temperature for ~20 mins to remove excess liquid.\n\n15.  Re-suspend the DNA pellet in 200 u1 of ddH2O and leave it at 4°C overnight \n\n16.  DNA concentration will be ~100ng /ul and can be determined using either an agarose gel or a spectrophotometer. \n\n \n\n                                                                            CTAB  DNA Isolation Buffer (500 ml) \n\n \n\n2% (w/v) CTAB (sigma)\n 10.0 g\n \n1.4M NaCI\n 40.91 g\n \n20 mM EDTA\n 20 m1 of 0.5 M EDTA \n \n100 mM Tris. HCI (pH 8.0)\n 50 ml of 1.0 M\n \nAdd ddH2O to make a final volume of 500 ml \n \n\n \n\nSDS Method \n\n1.      1-2 young leaves (~100mg) are placed in a labeled 1.1-ml collection tube or in a plate well, freeze-dried, and ground for 3-5 min at 30x/sec to a fine powder using a Mixer Mill and one 3-4mm steel bead in each well. (To isolate DNA from dry seeds, half to 3 seeds are wrapped with a weighing paper, crushed with pliers, placed in a col1ection tube or   in a deep-well plate ).\n\n2.      Preheat extraction buffer to 65°C in a water bath. (If the tissue samples are stored at -20°C, allow them to warm up to room temperature).\n\n3.      Add 0.2% (v/v) ß-mercap
182875
If a cell has a sequence of DNA that can be transcribed, that means it can be made into RNA. RNA is then translated into proteins for the cell to use. Every 3 RNA nucleotides codes for an amino acid which are used to build proteins. The proteins are built in the cell's ribosomes using messenger RNA and transfer RNA; both of which were transcribed from the original DNA sequence.\n\nSo the cell that has a sequence of DNA that can be transcribed has the ability to make protein structures for whatever it needs; enzymes or physical structure.
151519
Read as much as possible.  Also, University of Arkansas Press has a book "Writing Good Sentences" that is the best organized English Grammar workbook ever written.  Go to the college website to find it.  The author is a Dr. Faulkner, a professor there who invented the system while he was in the navy and the guys on the ship wanted him to teach them to write when they had spare time.
8687
mRNA  \nCapped messenger RNA is produced directly from your plasmid that contains a T7 or SP6 promoter. It is purified by our proprietary PUREmessenger™ technology. PUREmessenger™ is a chromatography that not only eliminates traces of DNA and proteins but also allows the selective recovery of the mRNA of the correct size. Shorter abortive transcripts as well as longer contaminating mRNA that are produced during in vitro transcription are eliminated. Capped mRNA will be purified by PUREmessenger™, quantified, re-suspended at 1mg/ml (except if otherwise specified) and aliquoted at 50 µl/aliquot (except if otherwise specified) in sterile conditions. It will be sent to you on cooling blocks together with its documentation: formaldehyde/agarose gel documentation picture. The capped mRNA that is sent to you ready-to-use (dissolved in water or the buffer of your choice) is 100% of your mRNA molecule of interest.\n\nCapped mRNA is produced in clean room facilities for research or clinical applications.\nYour benefit\nHighly purified, efficiently capped mRNA that has a superior efficacy compared to standard mRNA produced with in vitro transcription kits and simply purified by precipitation thus contaminated with alternative transcripts and traces of proteins and DNA fragments. The high quality of mRNA is reproduced from batch to batch. This guarantees the consistency of your results. Capped mRNA can be produced for research or for clinical utilisation: "documented" to be used as Active Pharmaceutical Ingredient or GMP-certified.
963400
well, i'm kind of a perfectionist when it comes to things like this...i make sure know my material, and have all my note cards in order. then, about a 1/2 - an hour beforehand i start doing deep breathing and stretching...it really works. practice your speech while looking in a mirror so you can see your facial expressions...practice reciting it to friends/family and get their feedback. while you're giving your speech, focus on ppl in the audience you know, and look at them from time-to-time. if you don't have to stand at a podium, move around a bit as you give your speech. try not to say "um" a lot...it makes it sound as if you don't know your material. try to smile at specific points in your speech material...if it's appropriate. that will make you look comfortable to your audience. i'm sure you will do a great job!! best of luck.
1020699
First of all make sure your template is good quality. Did you extract genomic DNA, do you have a library in plasmids...(?).\nTry re-purifying your template.\n\nMaybe your template is GC-rich or has some secondary structure.\nIs your template very long?If so make sure you let the polymerase is working at the elongating temperature for enough time (1min/kb)\n\nIf you are using plasmid template try not to use too much. Don't put more than 30-40ng DNA in a reaction. \n\nCheck your primers:\n-can they form a dimer?\n-can they anneal at more than one site on the template DNA with a only few mismatches?\n-is the difference in the Tm of the two primers more than 6 C (it shouldn't;ideally they should be the same and I try not to exceed a difference of 4)?\n-do your primers end in more than 3  c or g in a row?\n-what's their Tm and GC content?Tm should not be very low (I design my primers at 56-61) and GC 40-60%.\n\nThe most strigent conditions are using as annealing temperature 2 C below the lowest Tm of the two primers. Try lower temperatures.\n\nYou can try different enzymes Taq, Pfu, Pwo, etc, even mixtures. Not all enzymes have the same processivity.\n\nYou can try different Mg+2 concentrations.\n\nThere are a lot of control experiments you can do depending on your template and primers. Eg if you have a library in a  plasmid that has sites for universal primers you can use one such primer and the other respective primer you designed in order to check if there is something wrong with the quality of your primers. \n\nIf everything is perfectly designed and you've tried evreything but still get a very faint (but of correct size) band, gel purify it and use it as a template for a new PCR. In this case you'll have a very high chance of getting mutations so you'll need to use a high fidelity enzyme like Pwo or Pfu turbo and of course sequence a lot till you get a correct clone.
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Get radio time.  Send demos to major stations with some good PR.  It's even better if you can deliver them in person.\n\nThe best strategy is to have the best talent.  Harder done than said.