How to isolate DNA from seeds.?
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
697436
are you asking about DNA extraction or plant (apple tree) cross-breeding?
511827
Simply:\nTake sample cells from your body where a mutation is expected, and from a normal site.\n\nUse an enzyme to dissolve the cell's membrane\n\nUse alcohols to separate the DNA from the solution.\n\nTake the solution and use PCR (Polymerase Chain Reaction) to technically clone the DNA (make copies of it), but add in both normal and abnormal nucleotides (For both the mutated and non-mutated)\n\nPut it in a gel electrophoresis chamber and run electricity through the solution.\n\nYou can actually separate the abnormal nucleotides into specific chambers (A C G T )\n\nThe DNA fragments will separate throughout the gel and will create a pattern which shows the order of the nucleotides so you can find the specific code for the DNA.\n\nCompare the suspected mutated one to the normal cell.\n\n(It's pretty complex but that's the simple version)
32578
This is a fantastic question. There is an old joke that DNA extraction is like playing with test tubes of water. That's because DNA floating around in water is invisible, and looks like the researcher is just pouring clear water from one container to another.\n\nThere are several methods, called protocols, for isolating DNA from plants. A google search on plant dna extraction will give you a boatload of protocols. Most require exotic chemicals and lab equipment. \n\nSee the complicated lab method link below.\n\nBut you can extract DNA from plants in your kitchen using a blender, detergent, meat tenderizer, and rubbing alcohol! \n\nIn the case of Cleome, I would use about half a cup of the seed pods. Best of luck!
450846
I am  notsure if this is what you are looking for. also try the web site below\n\nTypical oil extraction from 100 kg. of oil seeds\nCastor Seed 50 kg\nCopra 62 kg\nCotton Seed 13 kg\nGroundnut Kernel 42 kg\nMustard 35 kg\nPalm Kernal 36 kg\nPalm Fruit 20 kg\nRapeseed 37 kg\nSesame 50 kg\nSoyabean 14 kg\nSunflower 32 k
122666
http://www.seedbiology.de/structure.asp
1002331
isolate the pathogen...
729453
K.C.'s answer works if you have a selective media that will work. Also (or in addition to that) you may want to plate out colonies and then PCR out a unique sequence or two.\n\nThis wouldn't be definitive but if you design your primers right it may work, but also may be very time consuming.\n\nAlso you may want to try restriction digests to remap your gene. This would work well, if you have a good map.\n\nGood luck, what a bummer
851316
I think I might be qualified to answer this question.\n\nDNA is separated in an electrophoretic field based on the size of the molecules.  \n\nImagine wet spaghetti strands (all different sizes) passing through a thick block of Swiss cheese.   Agarose electrophoresis is based on the ability of the smaller molecules to pass through holes easier than the larger molecules.\nThe smaller the mass, the more nimble it is, and the quicker it can manage all those "Swiss cheese" holes that agarose gels naturally have.\n\nOnce the DNA has run through the gel for a sufficient amount of time, you can stain the gel (actually you can stain the gel as the DNA moves through the gel too).  \n\nThe stain (the classic one is ethidium bromide, but there are others) has a propensity to squeeze itself between nucleotides on both strands.  When it is exposed to a transilluminator light  with a specific wavelength, the ethidium bromide will fluoresce and can be photographed for a permanent record.  \n\nAnalysis is typically based on running controls (positive and negative controls) and a size ladder along with the "unknown" PCR amplicon.  Usually I can predict the size of my unknown amplicon before I run the reaction. \n\nI can then choose a size ladder that will at least give me a rough idea of the size of my amplicon I expect. The ladder also allows me to observe any other false amplicons created during the PCR.  Additionally, some like to estimate how much PCR amplicon was generated in the reaction by comparing the intensity of the size ladder with the intensity of the amplicon. \n\nI hope I was able to help in answering your question.  Hoping YOU spread the good Karma!
381549
There are both culture-dependent and culture-independent ways of doing this...\n\nCulture-dependent: you can centrifuge a soil sample and then plate out the supernantant onto various types of agar.  If you are looking for bacteria, you can use agars that enrich for certain species like Streptomyces or Actinomyces Isolation Agar, or you can use more general agar like Tryptic Soy Agar.  Just be sure to include cyclohexamide in the media to prevent fungi from taking over!  If you are looking for viruses, you will have to do a virus/phage prep from the soil supernant and then plate onto lawns of bacteria or fungi, depending on what types of viruses you are looking for.  If you are looking for fungi, you can use the same supernatant but use media for fungi enrichment, such as Potato Dextrose Agar or Worts Agar.\n\nCulture-independent:  you can isolate DNA from the soil and do sequence analysis to see what kind of bacterial/viral/fungal commuunity there is in the soil.  There are various kits you can buy to do this, dependding on which organism's DNA you want.  This is a much faster method than growing them on agar, but you miss out actually seeing the organisms, as well as enumerating them.
376786
They get DNA from nucleated cells like white blood cells (in blood and saliva) and epithelial cells (in saliva). They lyze the cells to free the DNA, centrifuge and take the fraction with the DNA, treat and amplify or multiply the DNA fragments, put markers or specific labels, determine the presence and amount of this marked or labeled fragments, and compare the distribution with known samples or DNA database. DNA results are usually one page long.
530269
Stepwise-First you identify a gene of interest. Then, usually through years of work, you identify some function of the gene.  One way to identify this is to knock the gene out, a process called, approriately, gene knockout.  Knock out the gene in a cell line, or more interestingly, in an embryonic mouse- Now, if the embryo develops, you examine the animal, the tissue, the cellular funtions, and try to determine what has changed with the gene gone.  You can also overexpress the gene, so that it makes much more gene product, and see what happens then.  Once you have some handle on the function of the gene overall, you can make small mutations in it to determine exactly which part of the resultant protein does what.  Outside of scientific investigation, genetic engineering is used to make more viable or resistance plants (for example, put a gene that confers resistance to high salt, that you took from a plnat that grows in salty conditions, and put it into a strain of rice- now the rice can grow in high salt). It is also used to produce pharmaceuticals, such as antibodies- take a gene for an antibody out of the animal that makes it, put it in a bacteria or yeast, and then make tons of the stuff cheaply.
325792
of coarse. DNA is found in every cell. not just semen.
355375
Go here. http://www.dnacenter.com/\n\nYou can't make them get it done, but you can trick them into giving you DNA by collecting a straw they have used or a cigarette butt or, obviously, a semen sample. Anything with hair, skin cells, or spit will also work. There is no age or time limit.
514947
Everthing you ever wanted to know about DNA.\n\nhttp://www.dnaftb.org/dnaftb/
244250
You have to identify two sequences that are >6kb apart from one another that flank the sequence you are attempting to amplify. Better yet, you can get you a well understood cloning vector (plasmid) that already has primers develped for it that flank the polyrestriction site region (ie. puc19 [see link below], find the needed restriction enzyme sites that flank your desired 6kb sequence and clone the DNA into the vector. Be sure to check your sequence for any internal restriction sites. This may require you to do a partial digestion and recovery.
911470
how to make coffe from rice or okra seeds.\ntried and tested!!!