企業的存在靠的市場

企業的存在靠的市場

結果 (英文) 1:

The market on which the enterprise exists

結果 (英文) 2:

4. Material and Methods
4.1 Sample Collection
The protocol for capturing and sampling of bats in the present study was approved by the
Chung Yuan Christian University Animal Care and Use committee (approval number 103008)
and agriculture bureau of regional government. The captured bats were put into cotton bags
individually and allowed to produce fecal pellets for the collection in 500 μL of RNAlater®
RNA stabilization solution (Qiagen, Germantown, MD, USA). Blood samples were taken by
heparin (30 U/μL)-rinsed tips from the wing veins nicked by 26G needles. No more than 5%
of total blood volume of individual bat (1/20 of body weight) was collected for their survival
(Voigt CC and Cruz-neto A, 2009).
4.2 RNA Extraction
Viral RNA was extracted from the fecal pellets in RNAlater® solution by using QIAamp
Viral RNA kit (Qiagen). In brief, 140 μL of the supernatant of the RNAlater® solution with
fecal samples after the centrifugation at 5000 rpm for 20 minutes was added to 560 μL of
AVL buffer containing 5.6 μL of carrier RNA in AVE buffer. The mixture was incubated at
room temperature for 10 minutes and then added 560 μL of 96-100% ethanol. Next, 630 μL
of solution was applied to the QIAamp column in a 2-mL collection tube and centrifuged at
8000 rpm for 1 minute. The filtrate was discarded and the QIAamp column was washed by
AW1 and AW2 buffers. Finally, viral RNA was eluted from the QIAamp column in 60 μL of
AVE buffer after the centrifugation at 8000 rpm for 1 minute.
4.3 Reverse Transcription
The extracted RNA was reverse transcribed to cDNA by using iScript Select DNA Synthesis
kit (Bio-Rad, Hercules, CA, USA) containing the mixture of random primers and oligo dT
primers according to the manufacturer’s instructions. Briefly, the 20 μL-reaction comprising
13 μL of viral RNA, 4 μL of 5 x iScript reaction buffer, 2 μL of random and oligo dT primers,
6
and 1 μL of iScript reverse transcriptase was incubated at 25 oC for 5 minutes, 42 oC for 30
minutes, 85 oC for 5 minutes and then stopped at 4 oC.

結果 (英文) 3:

The existence of enterprises depends on the market