https://www.youtube.com/watch?v=GZejHrnWUXg /mnt/1T-5e7/mycodehtml/Biology/Gene_sequencing/Short_tutorial_text/Personal_Genomics_Institute/main.html ================================================================================ Decode: crypted gene -> decrypted gene ================================================================================ You convert "extracted DNA" into "DNA structure" where "decrypting DNA sequence" is "possible" ================================================================================ How to create "library" - There is an extracted DNA - Cut extracted DNA into the size you want to use by using ultrasound - DNA is randomly cut - Cut DNA - Manipulate structure of both ends of DNA to easily attach "adaptor molecules" - There are adaptors adaptors is the sequence which we already fully know adaptors is the label which is attached to unknown DNA sequence Attaching adaptor to DNA is called "ligation" ================================================================================ - There are many DNA pieces which are attached by adaptors ================================================================================ This means "you finish creating library" ================================================================================ Now, you should "decrypt" unknown DNA sequence of library cBOT machine: DNA amplication Flowcell: generates clusters Flowcell in zoom in Flowcell in more zoom in Flowcell is class plate which is coated with oligo ================================================================================ 8 oligos can take 8 kinds of DNA library? ================================================================================ You insert "flow cell" into "cBOT" machine to create "cluster" for DNA sequence decrypt ================================================================================ DNA library is flown into "flow cell" ================================================================================ Millions of library DNA are randomly attached to oligos ================================================================================ DNA library bends to next oligo ================================================================================ One more strand is generated by DNA polymerase creating 2 number of strands ================================================================================ ================================================================================ 2 strands again bend to next oligos ================================================================================ New strands are generated ================================================================================ Clusters which are composed of multiple single DNA piece library are generated ================================================================================ After creating cluster, you should perform "decryping (or decoding) unknown DNA sequence" ================================================================================ You insert "flow cell which has clusters" into gene analyzing machine ================================================================================ There are clusters ================================================================================ Primers (for decoding) are attached to adapter sequence - From this strand library, - Adapter sequence - Primer ================================================================================ Insert fluorescence labeled bases (A,T,C,G) and those bases will be attached to the unknown bases and take a picture ================================================================================ Blue colored base is attached to unknown base and take a picture ================================================================================ ================================================================================ ================================================================================ Decoded "unknow bases" are automatically stored into disk ================================================================================ Decoded DNA sequence 1st strand 2nd strand 3rd strand ... ================================================================================ From flow cell, one rane creates 2 files ================================================================================ ================================================================================ First file Second file ================================================================================ See the first one in detail ================================================================================ ================================================================================ - Sequence alignment - Align "decoded sequence" and "already previously decoded sequence" ================================================================================ Upper sequence: known decoded sequence Lower sequence: newly decoded sequence (short) Location of short piece of DNA can be confirmed ================================================================================ Find "variant" part ================================================================================