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allocation total RNK


Allocation RNK from thoraxes of bumblebees carried out with the help "trizolnogo" a method. For this purpose directed "trizolnyj" a reagent (38 % Phenolum, 0,8 M guanidin tiotsionat, 0,4 M of ammonium tiotsionat, 0,1 M sodium acetate, pH 5).

Homogenization and division of phases.
Inkubirovali navesku thoraxes of bumblebees (0,3) in 5 ml trizolnogo a reagent within 5 minutes at 60 s. Effected fabric homogenization in Potter's homogenizer within 1 minute. TSentrifugirovali the sample at 12000 g 10 mines at temperature 4 s. Received supernatant poured in other tube, a deposit rejected. Added 1 ml of chloroform in the tube and stirred on vortekse during 15 seconds TSentrifugirovali the sample 10000 g 15 mines at temperature 4 s.
Sedimentation RNK.
Carefully culled a water phase, deleted interfazu and the inferior phase. Added izopropanol and 0,8 M tsitrat sodium/1,2 of M NaCl Г2 of volume of a water phase. Tubes carefully stirred. TSentrifugirovali the sample at 10000 g 15 mines at temperature 4 s.
Flush RNK.
The received deposit was flushed by 7 ml of 74 % of ethanol. TSentrifugirovali the sample 10000 g 15 mines at temperature 4 s. The received deposit was dryed up by 5-10 minutes Added 80 mkl deionizirovannoj Н2О. A deposit resuspendirovali. Received RNK stored at - 20 s.
Concentration RNK defined by means of the spectrophotometer of Council of Federation-102 at length of a wave of 260 nanometers. Degree of cleanliness of the received preparations judged
On an interrelation And/and.
260 280
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A source: Syromjatnikov Michael Jurevich. BIOPOWER CHARACTERISTICS MITOHONDRIILETATELNYH of MUSCLES of BUMBLEBEES (BOMBUS TERRESTRIS L.). 2014

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