Wednesday, August 28, 2019

BIOCHEMISTRY CRITICAL REVIEW Essay Example | Topics and Well Written Essays - 750 words

BIOCHEMISTRY CRITICAL REVIEW - Essay Example The merit of RNA lies in its rapidity of expression (Kim and Eberwine, 2010).The main demerit of RNA is its transient nature; however, this may be controlled using stabilizers like StabiLizingUtr (Hayashi et al, 2010) but no such controlling factor was seen in the experimental protocol of Jankowska et al (2008) risking the expression period and genome integration (Kim and Eberwine, 2010). The transfection efficiency reported by Jankowska et al (2008) was about 30% using Lipofectamine 2000 transfection cationic liposome based reagent which is considered a highly efficient reagent (Dalby et al,2004) and has the capability of enhancing transfection efficiency in even poorly infectable cells such as CD34 (Byk et al,1998). Substitution of Lipofectamine2000 could have been done with 7.5% pegylated Lipofectamine which also produces high levels of expression (Decastro et al, 2006). The efficiency of transfection in the HeLa cells could have been increased using novel microfluidic sonoporation system which gives an efficiency of about 68.9% with 77% cell viability post insonication (Rodamporn et al, 2011). Verification of presence of hCG was done my immunohistochemical analyses using primary antibodies since immunohistochemistry is an important tool for biomarker detection (Luongo de Matos et al, 2010). Primary antibodies are used today for various analyses (Lipman, 2005). Detection was done in this experiment by using primary antibodies against hCG and the antigen-antibody complex was detected using flurophore, Cy3-conjugated antibodies. Primary rabbit polyclonal antibodies were used in the experimental protocol which was not absolutely specific for hCGÃŽ ² thereby increasing chances of non-specific binding. Polyclonal antibodies leads to cross-reactivity between divergent antigens (Frank, 2002).In the experimental protocol blocking buffer was used to reduce the non-specific binding and was

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