Name: qPCR Probes
Brand: Tsingke Biotech
SKU: 00302
Availability: InStock
Rating: 5 (1 reviews)

Advantages

Tsingke’s qPCR probes services offer numerous advantages, making them ideal for high-precision qPCR experiments. Our products, including custom qPCR probes and double-quenched probes, are synthesized in a state-of-the-art 100,000-level clean room, ensuring minimal contamination and high purity. With strict adherence to ISO 13485 standards, we guarantee custom yields qPCR probes with exceptionally low background noise, enhancing the reliability of results. Additionally, our expertise in qPCR probes design, combined with our ability to provide custom qPCR primers and qPCR primer and probes solutions, ensures optimal performance across diverse applications. Trusted by over 500 IVD companies, our qPCR probes synthesis process consistently delivers top-tier quality for both research and diagnostic use, making Tsingke a leader in customized qPCR probes.

Customer Trust

Serving over 500 IVD companies

Stable product quality

High-quality synthetic raw materials and technology, the probes have high purity and low background noise

Anti-contamination Procedures

Strictly control contamination from E.coli and human sources to avoid NTC peaks

Service Details

Tsingke provides a wide range of qPCR probes services, tailored to various scientific applications including qPCR probes design, custom qpcr probes, and qpcr primer and probes solutions. With lengths ranging from 15 to 40 nucleotides and options like MGB Probes, Molecular Beacons, and multiplex qPCR probes design, we ensure flexibility and precision for different research needs. The custom qpcr probes can be delivered as lyophilized DNA or in tubes, with options for purification methods such as DSL, OPC, PAGE, and HPLC. Our expertise in qpcr probes synthesis ensures high-quality results, which guarantee optimal performance for both research and diagnostics.

Service Name	Length (nt)	Purification	Price/ Turnaround time	Deliverable	Application
qPCR Probes	15-30	DSL/OPC/PAGE/HPLC/ Dual PAGE & HPLC	Inquire	· Tube or Customized · Lyophilized DNA ·COA Report (electronic)	The most commonly used types of qPCR experiments
MGB Probes	13-25				For qPCR experiments with higher TM
Double-Quenched Probes	15-45				For qPCR experiments with longer probes
Molecular Beacons	25-40				For qPCR experiments with extremely high sensitivity requirements
Other Probes	Customized	Customized			Special application directions

*Note: In addition to the recommended content, Oligo length and purification methods can also be customized.

Common Fluorescent Dyes and Quenchers types

Fluorescent Dyes	Max Abs	Max Em	Quenchers	Quenching range	Quenching Max
FAM	494 nm	518 nm	Dabcyl	380 nm-530 nm	452 nm
TET	521 nm	536 nm	Eclipse	390 nm-625 nm	522 nm
JOE	520 nm	548 nm	MGB	390 nm-625 nm	522 nm
VIC	538 nm	554 nm	TAMRA	470 nm-560 nm	544 nm
HEX	535 nm	556 nm	BHQ1	480 nm-580 nm	534 nm
Quasar 570	547 nm	570 nm	BHQ2	550 nm-650 nm	579 nm
Cy3	552 nm	570 nm	BHQ3	620 nm-730 nm	672 nm
TAMRA	565 nm	580 nm
ROX	585 nm	605 nm
Texas Red	595 nm	615 nm
Alexa Fluor 633	632 nm	647 nm
Cy5	643 nm	667 nm
Quasar 670	647 nm	667 nm
Cy5.5	684 nm	710 nm
Cy7	750 nm	773 nm

FAQ

What are the most commonly used fluorescent dyes?

FAM
TET
HEX
TAMRA
ROX
Cy3
Cy5

What causes low fluorescent signal in amplification curves?

First check the baseline-corrected or ROX-corrected raw curves. Low signal values in the amplification curves are mostly due to high background fluorescence. In probe-based detection, high background fluorescence is often caused by poor probe design, leading to insufficient quenching by the quencher group, improper pairing of the reporter and quencher groups, or low fluorescence labeling efficiency of the probe.

What causes erratic peaks in melting curves?

Erratic peaks in melting curves can result from contamination in the reaction system. Confirm contamination using NTC and NRC results, and check potential sources such as water, oligos, enzymes, and the experiment settings. Reagents exposed to strong light or high temperatures may degrade, so it is recommended to compare results with fresh reagents.

How to test the purity of primers?

A common method used in laboratories is the PAGE. For electrophoresis, use a polyacrylamide gel with 7 M urea, 20% polyacrylamide gel for primers with less than 12 bases, 16% polyacrylamide gel for primers with 12-60 bases, and 12% polyacrylamide gel for primers with more than 60 bases. Take 0.2-0.5 OD primer, dissolve with urea saturated solution or add urea dry powder into primer solution until saturated, and heat denaturation (95 ℃, 2 min) before loading sample. The purpose of adding urea is to denaturate, and to increase the specific gravity of the sample, which is easy to add the sample. After a certain period of time (about 2-3 hours), strip the polyacrylamide gel and detect the band type with a fluorescent TLC plate under the UV lamp. There is no impurity under the main band, indicating that the purity is good.
(Sometimes due to insufficient denaturation, there may be bands above the main band, which are primer secondary structure bands.)

Why do dissolved primers work fine at first but not after a period of time?

If the water in which you dissolve the primers has a low pH or is contaminated with bacteria or nucleases, the primers will degrade. Inadequate thawing and mixing during use, and uneven liquid may also cause inaccurate amounts of primers. It is recommended to aliquot primers, avoid repeated freezing and thawing, and use 10 mM Tris pH 7.5 buffer to dissolve primers. There is another possibility that there is no problem with the primers, but that the quality of the materials used in PCR, especially the template, is not completely consistent with that used previously.

Will the primers degrade when transported at room temperature?

It will not degrade, and the dry primer can be stably stored at room temperature for at least two weeks. The average transport time is usually 5days, so the primers you receive will not degrade.

How to measure the OD value of primers?

The OD value of the synthetic primer was determined by using an ultraviolet spectrophotometer with a wavelength of 260 nm and a quartz colorimetric cup with an optical path of 1 cm to determine the optical density of the solution. The optical density of the solution should be diluted to 0.2-0.8. After the DNA dry powder is fully oscillated and dissolved with a certain volume of water, the OD value is measured by diluting with 1 mL of water. OD value of mother liquor should be converted according to dilution ratio. For example, to verify whether the amount of 2 OD primer is accurate, the simple practice is to add 1 mL water, thoroughly dissolve and mix, take 100 μL, add 900 μL water, a quartz colorimetric cup with a light diameter of 1 cm, a wavelength of 260 nm, and the absorbed reading at this time is 0.2.

How many primers of OD value should be synthesized?

According to the purpose of the experiment. In general, about 20 bases of primer 2 OD in PCR amplification can do 400 rxns standard PCR reactions of 50 μL system. If it is to do gene splicing or annealing to do the ligation, 1 OD is enough.