线上葡京电玩

Design your assay in three easy steps

线上葡京电玩

PRODUCT DISCONTINUATION NOTICE

Please be informed that The Universal Probe Library 葡京国际在线官网 Center will be accessible with limited availability until December 31, 2020. Should you have any questions regarding the 葡京国际在线官网 Center or availability of the Universal Probe Library products, please contact Merck KGaA technical support for further information.

Based on only 165 short hydrolysis probes substituted with Locked Nucleic Acids, 线上葡京电玩 (UPL) allows you to design real-time qPCR assays in seconds and analyze over five million transcripts of virtually any sequenced organism. The easy-to-use ProbeFinder 葡京国际在线官网 Software displays the target-specific primer sequences (ready for ordering at your preferred oligo supplier) and the matching 线上葡京电玩 probe.

线上葡京电玩 assays are compatible with all real-time PCR instruments capable of detecting fluorescein, FITC, FAM, and/or SYBR? Green I, and follow standard cycling protocols for hydrolysis probe assays.

Choose the different 线上葡京电玩 Products:

PRODUCT DISCONTINUATION NOTICE

Please be informed that The Universal Probe Library 葡京国际在线官网 Center will be accessible with limited availability until December 31, 2020. Should you have any questions regarding the 葡京国际在线官网 Center or availability of the Universal Probe Library products, please contact Merck KGaA technical support for further information.

线上葡京电玩 System Technology

LNA-based Probes

UPL is based on only 165 short hydrolysis probes. They are labeled at the 5' end with fluorescein (FAM) and at the 3' end with a dark quencher dye.

The extensive transcript coverage of the UPL probes is due to their short length of just 8 - 9 nucleotides and the selected sequences. In order to maintain the specificity and melting temperature (Tm) that hybridizing qPCR probes require, Locked Nucleic Acids (LNA) are incorporated into the sequence of each UPL probe. LNA's are DNA nucleotide analogues with increased binding strengths compared to standard DNA nucleotides.

The sequences of the 165 UPL probes have been carefully selected to detect 8- and 9-mer motifs that are very prevalent in the transcriptomes, ensuring optimal coverage of all transcripts in a given transcriptome.

Frequency of 9-mer occurrence

Within the human transcriptome, each probe binds to approximately 7,000 transcripts, while each transcript is detected by approximately 16 different probes. Only one specific transcript is detected at a time in a given PCR assay, as defined by the set of chosen PCR primers. For each assay, the design software suggests an optimal set of PCR primers, a probe, and any possible alternative sets.

UPL assays are compatible with all real-time PCR instruments capable of detecting fluorescein, FITC, FAM, and/or SYBR? Green I. UPL assays have been used successfully on the LightCycler??480 System, the LightCycler??Carousel-Based System, and other real-time PCR instruments from several suppliers.

Read more about?线上葡京电玩 System Performance Data

LNA Technology

Comparison of LNA, DNA, and RNA.

Locked Nucleic Acids (LNA) is a class of nucleic acids analogues, where the ribose ring is "locked" with a methylene bridge connecting the 2'-O atom with the 4'-C atom. LNA nucleosides containing the six common nucleobases (T, C, G, A, U and mC) that appear in DNA and RNA are able to form base-pairs with their complementary nucleosides according to the standard Watson-Crick base pairing rules.

The Figure shows the comparison of LNA, DNA, and RNA.

Comparison of mismatch conditions for LNA and DNA

Therefore, LNA nucleotides can be mixed with DNA or RNA bases in the oligonucleotide whenever desired. The locked ribose conformation enhances base stacking and backbone pre-organization, this gives rise to an increased thermal stability and discriminative power of duplexes. LNA discriminates single base mismatches under conditions not possible with other nucleic acids.

The Figure shows the comparison of mismatch conditions for LNA and DNA.

Multiplex 澳门葡京备用网址

Multiplex 澳门葡京备用网址 with 线上葡京电玩 Reference Gene 澳门葡京备用网址

Use the 线上葡京电玩 Reference Gene 澳门葡京备用网址 together with the 线上葡京电玩 to easily quantify expression levels of a human, mouse, or rat gene of interest in relation to an endogenous reference gene in a dual-color assay. Three reference gene assays are available for human (G6PDH, GAPD and ACTB- gene), 2 for mouse (ACTB-, and GAPD gene each).

Each 线上葡京电玩 Reference Gene Assay provides a specifically designed 12-mer UPL reference gene probe and the corresponding reference gene-specific primer pair in a separate tube. The probe is labeled with 葡京线上游戏appYellow 555 at the 5? end and with a dark quencher dye near the 3? end, to enable dual-color assays together with the standard UPL probes, which are labeled with FAM. The UPL reference gene probes can be detected using real-time PCR instruments with excitation filters of 470 nm to 530 nm and emission filters of 550 nm to 610 nm. In multiplex assays, a standard FAM or SYBR? Green I filter is used as the first channel to detect FAM-labeled UPL probes for the gene of interest, and a longer-wavelength emission filter (usually the next possible emission filter moving to longer wavelengths, e.g., 560 nm or 568 nm) is used as the second channel to detect UPL reference gene probes.

Multiplex PCR assays for a target gene and a UPL Reference Gene are designed by the ProbeFinder software at the 葡京国际在线官网 Center with a high success rate of over 90%. In case no matching assays are found for the reference gene of choice (out of the listed assays), ProbeFinder SW suggests dual-color assays with one or more other reference genes out of the list.

Designing multiplex assays is as easy as designing monoplex assays: when you select the multiplex option online, ProbeFinder will design multiple UPL assays for your gene of interest. At the same time, each assay is subjected to in silico PCR testing to verify that it can be multiplexed with one (or more) of the UPL Reference Gene 澳门葡京备用网址 for the relevant organism.

Expression of 45 genes detected with a 线上葡京电玩 dual color assay

Using the LightCycler??480 澳门葡京地址 System, the expression of 45 genes is detected with a 线上葡京电玩 dual color assay when testing only one primer and probe combination (examples are shown in Figure right). The detected Cp values of dual-color assays differed by no more than 0 to 2.0 cycles from those detected with the single-color approach.

The Figure shows the expression of 45 genes detected with a 线上葡京电玩 dual color assay (Data kindly provided by Professor Stephan Wiemann, German Cancer Research Center, Heidelberg, Germany).

For more detailed description:

线上葡京电玩 Brochure

ProbeFinder Quick Reference for 葡京国际在线官网

Click here to read about complete, function-tested 葡京线上电子 RT-qPCR assays for human, mouse, and rat targets of your choice, based on UPL technology.

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