Electrophoresis analysis of PCR products rapidly

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analysis of PCR products

Fast electrophoresis analysis of PCR products

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Analysis your PCR products in only 6 minutes                   

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How to analysis your PCR products in only 6 minutes?

PCR and gel electrophoresis are two steps of PCR technique. In a protocol of PCR gel analysis, PCR products were loaded into gel wells

Agarose has been widely used for PCR gel electrophoresis in DNA RNA analysis procedures because agarose powder can be easily  heated to form an agarose gel with sample wells. In PCR gel electrophoresis method, for example, a microwave of 1 minute heating can melt agarose powder into buffer solution. The agarose gel solution is then cooled down to form an agarose gel.

The Problem of your old PCR gel system

1x running buffer, a highly conductive liquid, has two functions in conventional submarine agarose gel electrophoresis: keeping the agarose gel cooled and conducting electric current.

horizontal gel electrophoresis diagram

To avoid gel overheating, the volume of running buffer should be as large as possible. But more buffer conducts more electric current, which generates more heat. So buffer volume should be as small as possible. Which is correct - more buffer or less buffer?

 

  Breakthrough in rapid electrophoresis with no running buffer

  See User Feedback about the performance of buffer-less high speed electrophoresis:

" The Liberty bufferless gel system is the best gel electrophoresis system we've ever used. It's not only easy to set up, robust, and fast, but generates excellent results without the need of running buffer. Highly recommended. See our genotyping service picture below." ---- Ivan Delgado Orlic, President and CEO, Mouse Genotype Co. 

 

 

The principle of the 6Mgel invention

The inventor of 6mgel recognized that water is the best and cheapest coolant. The electrical resistance of lab dd-water is 18MΩ, which means that dd-water is an excellent electrical insulator. By simply adding dd-water directly on top of the buffer-containing agarose gel, the desired effects are achieved: gels can be run much faster!  You can add a large volume of dd-water to dissipate heat so that a much higher voltage can be applied to drive faster migration of DNA molecules. Band "smiling" is eliminated because the large volume of dd-water in direct contact with the gel maintains even heat distribution. Also, sharper banding occurs due to "stacking effects of dd-water in gel wells.

In 6Mgel, the V/cm in wells is elevated about 3-fold higher than in the gel body, which drives DNA 3 times faster in wells than in the gel body, resulting in sharper bands. Furthermore, consumption of running buffer can be reduced by 50-75%.

 

During the development of 6Mgel, more and more features were added, such as buffer-less high speed electrophoresis, lab-made precast gels for high-speed electrophoresis, and agarose-saving gel systems. Go to 6mgel systems for more information.

Q & A:

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What kind of agarose grade is required for high speed PCR electrophoresis: ---- In PCR electrophoresis, agarose quality is important. But our high speed electrophoresis technique was designed for any grade of agarose powder on the market. So far, over 86 different kinds of agarose have been used successfully in our Liberty 120 gel system for rapid electrophoresis.

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How electrophoretic power is connected: ---- PCR gel is usually immersed under electrophoresis buffer for electrophoretic connection. In our techniques, The body of PCR gel is immersed in cooling water. Two ends of PCR gel are exposed to buffer reservoirs for electrophoretic connection. This setup reduces temperature of PCR gel during electrophoresis so that high voltage can be used to drive DNA samples with rapid electrophoretic migration.

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What kind of electrophoresis buffer used for high speed PCR electrophoresis: ---- For PCR electrophoresis, 1X TAE buffer, 1x TBE buffer are the most popular buffers of electrophoresis, For denaturing electrophoresis, MOPS is the choice of buffer in high speed electrophoresis.

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Why Liberty 120 is the best gel system for high concentration PCR electrophoresis: ---- First, high resolution agarose is the most expensive agarose on the market, usually over $3 for each gram of agarose. Second, high resolution agarose is used at a very high concentration which make an PCR gel extra costly. Third, high concentration agarose makes electrophoretic migration of DNA very slow in conventional electrophoresis, usually over hours. With Liberty 120 gel system, you can save agarose with 3 gel sizes, you can run gels at much higher voltage to generate powerful electrophoretic force. The costs of time and agarose are reduced.

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Why PCR gel can be immersed in cooling water: ---- Agarose solution in TAE buffer forms a solid agarose gel when cold down in gel tray. By soaking the PCR gel in a cooling water without running electrophoresis, the buffer ions in PCR gel body can release slowly from PCR gel body to cooling water. But, during high voltage PCR electrophoresis, ions are driven by powerful electrophoretic force between two electrodes and move horizontally across PCR gel body which prevents the escape of ions from PCR gel to cooling water.

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Why high speed PCR gel electrophoresis shows straight banding: ---- The rate of electrophoretic migration of DNA through PCR gel is depend on voltage, agarose concentration, and temperature. In regarding to the issue of "band smiling", the concentration of agarose is uniform and can be removed from our discussion. For factor of temperature, in traditional PCR electrophoresis, PCR gel is immersed under a thin layer of electrophoresis buffer (the layer of buffer has to be thin for reducing electric current). The cooling function of the buffer liquid is very weak. The local temperature across sample lanes of a PCR gel during electrophoresis varies significantly which is the main source of "gel band smiling". In Liberty 120 gel system, agarose gel is immersed under large volume of cooling water, every points of the entire PCR gel remain in uniform temperature during electrophoresis, this is, gel smiling is substantially removed.

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Can we run sybr green pcr high speed --- Yes. Sybr green pcr samples show the same bands as regular tagman pcrs.

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Can we see pcr primers on high speed gel --- The length of PCR primers are usually 18-30 bases. Frequently, PCR primers form dimers in gel electrophoresis. Therefore, when running a high concentration gel, such as 3% high resolution agarose gel, we can see PCR primers in two bands. The first band is primer in single formate. The second band is primer dimer.

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What are the differences between PCR and real time QPCR --- When compare the real time PCR with regular pcr amplification methods, the key point are: A. The regular taqman PCR gives your the final result of the amplification. But real time PCR shows you the speed of template detection and amplification. That is, Real time Q PCR offers you a quantitative measurement of your template. Higher quantity of template reaches maximum amplification in earlier cycles and lower quantity of template reaches maximum amplications in delayed cycles of real time pcr analysis. B. In real time q pcr, 3 primers are required. The 3rd primer is a probe to report the progress of the real time pcr in each thermal cycle. That is why it called real time q PCR. While regular PCR provides no information about the thermal cycles.

6Mgel Innovative Gel Tips

What is 6Mgel Confidence?

6Mgel has been offering Guaranteed Return to all customers for 16 years. Any gel equipments can be returned for any reason without rejection. 6Mgel quality shows a total of 2 unit being returned and 4 units being repaired among all units sold.

 

How to electrophoresis PCR products?

How to run high-speed gel with no running buffer

Hot to eliminate agarose gel band smiling?

How to save agarose?

Explore the 6Mgel website --

You will find techniques and tips that can help you produce data more quickly and easily, with exceptional results. 

 

 

 

 

 

 

 

Liberty 120 high- speed gel system

The most recognized 6Mgel model

See Genotyping / PCR results in 6 minutes

 

Problem of old PCR gel electrophoresis:

The agarose gel formation is readily reversible by heat. Over heating will melt the agarose gel. That is the reason why gel electrophoresis of PCR samples in most labs were performed slowly at low voltage.

The breakthrough of high speed PCR gel electrophoresis in 6 minutes:

By designs of reducing heat generation and cooling enhancement, Biokeystone overcomes the problem of PCR gel melting in electrophoresis. Higher power drives DNA samples run through PCR gel in just 6 minutes without over heating the agarose gel.

To know more PCR product of gel electrophoresis, please visit the website and email to info@6mgel.com for user manuals.