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picture1_Hybridization Staining


 180x       Filetype PPT       File size 0.94 MB       Source: uvm.edu


File: Hybridization Staining
the following hybridization mix is prepared for each sample fragmented crna 5ug 10 ul control b2 oligo 1 7 ul 20x eukaryotic control mix 5 ul herring sperm dna 1 ...

icon picture PPT Filetype Power Point PPT | Posted on 03 Sep 2022 | 3 years ago
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       The following hybridization mix is prepared for each sample
      Fragmented cRNA              5ug                                          10 ul       
       Control B2 Oligo                                                         1.7 ul
      20x Eukaryotic Control mix [bio B, bio C, bio D, Cre]                      5 ul 
      Herring Sperm DNA [10mg/ml]                                                1 ul 
      Acetyleted BSA [50mg/ml]                                                   1 ul
      DMSO                                                                      10 ul
      2x Hybridization Buffer                                                   50 ul
      Water                                                                     22.3 ul
                                          Denature 99C               Inject into
                                          10 minutes                 GeneChip
                                                       
             The hybridization oven
                Target binds to the Probes
                              
              RNA-DNA Hybridization
    Targets:
    Antisense 
    biotinylated
     cRNA
       Probe sets: The DNA oligo probe is attached to the 
                    GeneChip via a silane bond
                                  
                                  Hybridization
         Optimized Hybridization is the process of single stranded nucleic acids binding 
               to another strand with identically complement sequence [We hope] 
                                      Types:   DNA to DNA     
                                                DNA to RNA 
                                                RNA to RNA 
                                                PNA to DNA 
                                                              
                                              
                                            
                    Stringency  
     Stringency is a condition that causes a change in the local 
    hybridization environment and “interferes” with the binding kinetics
    Stringency prevents: 
    .    Binding of non-complementary strands 
         Self hybridization – hairpin formation
         Disassociation of strands
                           
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...The following hybridization mix is prepared for each sample fragmented crna ug ul control b oligo x eukaryotic herring sperm dna acetyleted bsa dmso buffer water denature c inject into minutes genechip oven target binds to probes rna targets antisense biotinylated probe sets attached via a silane bond optimized process of single stranded nucleic acids binding another strand with identically complement sequence types pna stringency condition that causes change in local environment and interferes kinetics prevents non complementary strands self hairpin formation disassociation...

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