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Shotgun Cloning Method ExplainedShotgun Cloning Method Explained Shotgun Cloning Method Explained


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Shotgun sequencing method

Shotgun DNA sequencing Topic index 1) principle 2) procedure a) DNA fragmentation b) Agarose gel electrophoresis c) purification and modification d) cloning e) sequencing f) overlapping 3) Advantage 4) Limitations Fragmentation Fragmentation of the DNA can be done by using restriction enzymes, physically by breaking it into small pieces (usually 2, 10, 50, and 150 kb ) by passing it through a narrow gauge syringe or sonicating it, which is the way of breaking the sample using sound waves, by mechanical method, shearing It is a random process, so the sequences of the fragments will have some overlap between them Fragments of about 150Mb are obtained Cloning Next step is joining of DNA fragments with a vector which is a carrier DNA This method is known as cloning And a sequence library is created Sequencing of entire genome will be created by using this library Sequencing Sequencing of each clone in library is done Individual fragments are sequenced individually using the chain termination method to obtain reads Multiple overlapping reads are obtained for the target DNA by performing several rounds of this fragmentation and sequencing Advantages and Disadvantages of Shotgun Sequencing Shotgun sequencing had a number of important advantages over previous methods 1)Faster because the mapping process was eliminated 2)Uses less DNA than other methods 3)Less expensive than approaches requiring a map Some disadvantages of shotgun sequencing include 1) Requires computer processing power beyond what an ordinary laboratory would possess 2) Can introduce errors in the assembly process 3) Requires a reference genome 4) May not be able to assemble repetitive sequ

Shotgun Gauges Explained Shotguns 101 #3

The shotgun world is a mess of old standards & notations, and nowhere is this more evident than the way we define bore diameters Just about everyone is familiar with terms such as 12-gauge or 20-gauge, but what exactly does gauge mean, and how does it translate to an actual diameter? This video explains the history and math behind the gauge system used to define shotgun bores, and discusses the shotshell sizes in use today Please note that any comments on the relative popularity and availability of the shotshell sizes and loads discussed here are from a North American perspective Things may be different in other parts of the world The Shotguns 101 video series focuses on general shotgun topics I already have a lot of ideas for future videos, but I d still like to hear any suggestions you might have I recommend watching this video with the annotations turned on I ll be adding various notes to provide further details and address some common questions and potential points of confusion There will be no giant flags blocking the video, and absolutely no begging for sub ions or likes Shotguns 101 Playlist s www youtube com playlist?list PLzPVXnz7gkEWz4JXaAQ7zEeWVBsaN4c3K Remington 870 Tips & Tricks Playlist s www youtube com playlist?list PLzPVXnz7gkEXSPRg6bGdbquEwfJFupUYL PROGRAMS USED Adobe Reader 9 Audacity AVS Video Editor 6 3 Google Chrome Microsoft Excel Microsoft PowerPoint Microsoft Word Rhinoceros 3 0 Windows Paint Windows Picture and Fax Viewer DISCLAIMERS By accessing this video, the viewer signifies their understanding of, consent to, and agreement with the following All products and copyrighted works mentioned or shown herein remain the property of their respective creators and or owners I make no claim to anything in this video that I didn t personally create Any such elements are used herein for educational and informative purposes; and the purpose, character, and substantiality of their use does not, to the best of my understanding, constitute copyright infringement according to 17 USC §

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