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2.2.3.2. Allocation total RNK from grudok bumblebees and reverse transcription


ekstraktsiju total RNK from thoraxes Bombus terrestris carried out "trizolnym" a method. Qualities of a preparation received RNK checked with the help elketroforeza (fig. 22).



Fig. 22. elektroforegramma total RNK from thoraxes B. terrestris.
On given elektroforegramme it is visible, that quantity 28S rrnk above, than 18S rrnk it means that there was no degradation RNK under the influence of RNKaz. Also there is no DNA admixing. Thus, given sample RNK approaches for the further operations (reverse transcription and then PCR-reaction).
For synthesis kdnk the anatropic transcriptase of virus Molony lejkoza mice M-MuLv was used. Enzyme shows 5 ’ ^3 ’ polimeraznuju activity, effective only concerning templates RNK. In quality zatravochnogo prajmera it was used oligo - (bt) 2о prajmer. Use
prajmerov the given phylum allows to synthesise moleuly kdnk exclusively from mrnk. During reverse transcription the wide panel of products of DNA of various length (fig. 23 is formed.). Received kdnk used for PCR-reactions.


Fig. 23. elektroforegramma kdnk from thoraxes B. terrestris.

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A source: Syromjatnikov Michael Jurevich. BIOPOWER CHARACTERISTICS MITOHONDRIILETATELNYH of MUSCLES of BUMBLEBEES (BOMBUS TERRESTRIS L.). 2014

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