Biology Reference
In-Depth Information
9. The amount of extract required may vary depending on the
luciferase expression level and the instrumentation used; the
amount used should be adjusted to keep the signal within the lin-
ear range of the assay.
10. At low pH the wavelength of emitted light from the FF reac-
tion is shifted to lower wavelengths.
11. Control of temperature is also important since the use of non-
equilibrated reagents directly from the refrigerator can cause a
5-10 % decrease in efficiency.
12. Depending on the luminometer used it might be necessary to
prime auto-injector systems with buffers. Calculate additional
volume of the buffers for this.
References
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a dual luciferase reporter pull-down assay sys-
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2. Allegra D, Mertens D (2011) In-vivo quantifi-
cation of primary microRNA processing by
Drosha with a luciferase based system. Biochem
Biophys Res Commun 406:501-505
3. Jin Y, Chen Z, Liu X et al (2013) Evaluating
the MicroRNA targeting sites by luciferase
reporter gene assay. Methods Mol Biol 936:
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4. Bronstein I (1994) Chemiluminescent and
bioluminescent reporter gene assays. Anal
Biochem 169-181
5. Stables J, Scott S, Brown S et al (1999)
Development of a dual glow-signal, firefly and
renilla luciferase assay reagent for the analysis
of G-protein coupled receptor signalling. J
Recept Signal Transduct Res 19:395-410
6. Winter J, Jung S, Keller S et al (2009) Many roads
to maturity: microRNA biogenesis pathways and
their regulation. Nat Cell Biol 11:228-234
7. Zeng Y, Yi R, Cullen BR (2005) Recognition
and cleavage of primary microRNA precursors
by the nuclear processing enzyme Drosha.
EMBO J 24:138-148
8. Han J, Lee Y, Yeom K-H et al (2006) Molecular
basis for the recognition of primary microRNAs by
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