Black Hole Spy 0.5
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Black Hole Spy 0.5
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A slow orb of void that eats through all obstacles
Destroys all Materials caught in its radius.
A slow orb of void that eats through all obstacles and casts another spell as it expires
Destroys all Materials caught in its radius.
Categories :
Limited use spells
Spells with positive cast delay
Spells
Projectile spells
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Black Hole is a somewhat uncommon projectile Spell which summons a dark orb that travels slowly, attracting objects and consuming any terrain in its path. Despite its similarity to Giga Black Hole , this more contained version does no damage to enemies or the player.
It also comes as a variant with an expiration / death trigger, which behaves the same way, but costs 20 additional mana and will cast the next spell when the black hole expires. This can be used to "chain" black holes together, to create longer tunnels, in the absence of a more convenient modifier like Speed Up .
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rhAmp SNP Genotyping
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Selecting the correct reporter dye and quencher Reporter dyes: The correct reporter dye will depend on the type of instrument you are using and
the compatibility of the dye with the instrument. Please see the instrument compatibility table for a list of reporter
dyes compatible with common instrumentation. For multiplexing applications, we recommend using reporter dyes with
the least amount of spectral overlap. For assistance in selecting multiplex dye combinations, use our Multiplexing Dye Selection Tool , which helps you select dyes based on what instrument you have. As an additional reference, for
a list of selected IDT dyes and quenchers, please see the dye and quencher wavelength figure and the instrument compatibility
table.
Quenchers: Traditional dark quenchers absorb broadly and do not emit light, which allows for the
use of multiple reporter dyes with a given quencher. This characteristic allows for expanded options for multiplex
assays. Dark quenchers simplify detection, which makes them compatible with a broad range of image analysis instruments. These double-quenched probes are an improvement over traditional dual-labeled probes and have consistently
low background, reduced C q values, improved precision, and enable the use of longer probes for design in AT-rich
regions. Double-quenched probes are available with a variety of fluorophores, including SUN, FAM, HEX, TET, Yakima Yellow, and Cy 5. For more information download the PrimeTime Custom qPCR Probes flyer. We supply commonly used dark quenchers
as well as the proprietary dark quenchers, Iowa Black FQ, Iowa Black RQ, and the internal ZEN Quencher. TAMRA is
also a quencher option for a FAM reporter dye.
Summary tables:
Instrument compatibility table Dye and quencher wavelength chart
Which dyes are available for use with the IDT ZEN quencher?
How do I dilute and store PrimeTime qPCR Probes?
SUN fluorophore—a molecular equivalent to VIC »
qPCR Probes—selecting the best reporter dye and quencher »
+32 (0) 16 28 22 60 eutechsupport@idtdna.com
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Generate reliable, high-quality gene expression analysis data with double- and single-quenched fluorescent hydrolysis probes.
PrimeTime™ qPCR Probes are 5′ nuclease probes, available with an assortment of reporter-dye combinations that are compatible with common qPCR instruments and that include several license-free combinations.
* With the exception of mixed base oligos, which could potentially represent multiple sequences and therefore cannot be accurately evaluated by ESI mass spectrometry.
Yakima Yellow (YAK) is a registered trademark of EliTech Group, Cy is a registered trademark of Cytiva, and Texas Red is a registered trademark of Molecular Probes, Inc.
Choose from a wide variety of synthesis scales, dyes, and quenchers, including several license-free options. Prices and estimated turnaround times depend on the degree of complexity. For more information on dyes and quenchers, see Product details. Guides
for compatible dye and quencher combinations are available in the Resources tab. For multiplex applications, consult the PrimeTime Multiplex Dye Selection tool.
The estimated turnaround time for standard level probes is 3–5 business days. The minimum guarantees are listed for probes that are 15–35 bases in length.
* ZEN/Iowa Black™ FQ is a double-quenched probe, which provides superior performance compared to traditional single-quenched probes. For more information, download the PrimeTime Custom qPCR Probes Flyer .
The estimated turnaround time for complex level probes is 5–7 business days. The minimum guarantees are listed for probes of 15–35 bases.
* ZEN/Iowa Black FQ is a double-quenched probe, which provides superior performance compared to traditional single-quenched probes. For more information, download the PrimeTime Custom qPCR Probes Flyer .
TEX™ fluorophore and JOE are trademarsk of ThermoFisher; TYE™ is licensed from ThermoFisher; Texas Red ® is a registered trademark of Molecular Probes, Inc.
Available with a 5′ FAM and the option of a 3′ IBFQ quencher alone or in combination with the internal ZEN quencher. Estimated time to shipment is 3–5 days.
HPLC purified. Availabile for lengths of 18–35 bases. Shipped dry. Minimum guarantee of 5 nmol.
Additional shipping and handling fees for expedited service apply.
* ZEN/Iowa Black™ FQ is a double-quenched probe, which provides superior performance compared to traditional single-quenched probes. For more information, download the PrimeTime Custom qPCR Probes Flyer .
While traditional probes have approximately 20–30 bases between the fluorophore and the quencher, the internal ZEN or TAO Quencher decreases that length to only 9 bases (Figure 1). This shortened distance, particularly when combined with the traditional
3′ end quencher, leads to a much more thorough quenching with much lower background and enables the use of much longer probes for designing in AT-rich target regions. In addition to the significantly decreased background, Double-Quenched Probes
also provide consistently reduced C q values and improved precision when compared to traditional probes. Use of Double-Quenched Probes can allow you to experience both increased sensitivity and precision in their qPCR experiments.
Figure 1. Double-quenched probe showing position of internal ZEN Quencher.
For more information download the PrimeTime Custom qPCR Probes Flyer .
The smaller scale and lower price of PrimeTime Mini qPCR probes make them ideal for digital PCR (dPCR) applications, testing a new probe for gene expression or genotyping applications, or for testing the conversion to FAM/IBFQ and FAM/ZEN/IBFQ from other FAM-related quenchers. Mini
probes are offered with 5′ FAM, SUN, HEX, and Cy5 fluorophores. Quenchers include 3′ IBFQ alone, double-quenched internal ZEN with 3' IBFQ, or double-quenched internal TAO with 3' IBRQ (Cy5 only).
The PrimeTime Eco qPCR Probe is the ideal scale when you need to perform ~500 reactions for gene expression analysis or genotyping. The combination of medium scale and low cost is ideal for initial screening of large sample sets. Eco scales are also amenable to dPCR applications. Eco qPCR Probes are available as double-quenched probes with 5′ FAM, SUN, HEX, and Cy5 fluorophores. Quenchers include 3′ IBFQ alone, double-quenched internal ZEN with 3' IBFQ, or double-quenched internal TAO with 3' IBRQ (Cy5 only).
Express qPCR Probes are HPLC-purified qPCR probes that are assessed by mass spectrometry for quality control, with traces provided on our website free of charge*. Due to the turnaround time, these probes have
limited dye-quencher combinations and synthesis scales.
* With the exception of mixed base oligos, which could potentially represent multiple sequences and therefore cannot be accurately evaluated by ESI mass spectrometry.
Step 1 —The primers and probe hybridize in a sequence-dependent manner to the complementary DNA strand. Because the probe is intact, the fluorophore and quencher are in close proximity and the quencher absorbs fluorescence emitted by the fluorophore.
Step 2 —The polymerase extends from the primers and begins DNA synthesis.
Step 3 —The polymerase reaches the probe and the exonuclease activity of the polymerase cleaves the hybridized probe. As a result of cleavage, the fluorophore is separated from the quencher and fluoresces.
Step 4 —The fluorescence is detected by the real time instrument.
These steps are repeated for each PCR cycle and allow detection of specific products. When using intercalating dyes, such as SYBR ® Green I (Life Technologies, Inc.), primer-dimers and non-specific products will also contribute to fluorescence. In contrast, the 5′ nuclease assay is specific and fluorescence will only be detected for the DNA sequence to which the probe and primers hybridize.
To demonstrate the performance of different dye-quencher combinations, we tested a dilution series and found robustness in PCR efficiency and R 2 values across all dye-quencher combinations available (Figure 1).
Figure 1. Demonstrated assay performance with multiple dye–quencher combinations. A plasmid dilution series of the CSK (c-src tyrosine kinase) Assay was used to test different dye-quencher combinations. The data illustrates robustness in PCR efficiency and R 2 values close to one across all dye/quenchers available for PrimeTime Assays. All reactions were run with TaqMan ® Gene Expression Master Mix (Thermo Fisher) under standard cycling conditions. The first four assays were run on the 7900 Fast Real-Time PCR System (Thermo Fisher) and the final assay (Cy5) was run on the LightCycler ® 480 System (Roche).
Figure 2. PrimeTime™ SUN-labeled probes deliver comparable performance to VIC/MGB-NFQ probes. A double-quenched SUN/ZEN/IBFQ probe was compared to a VIC ® /MGB-NFQ probe in a qPCR assay for GUSB and PGK1 .
gBlocks™ Gene Fragments with sequences identical to GUSB and PGK1 were analyzed over 6 sequential 10‑fold dilutions (10 2 –10 7 copies). Normalized fluorescence values were plotted (A) including
and (B) subtracting background fluorescence. C q values were comparable, and the final R 2 v
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