The INGENYphorU is a vertical electrophoresis instrument based on Ingeny’s proprietary U-shaped spacer. This spacer design allows 100% leak-free casting of all types of gels; gels are cast directly in the instrument. Casting a gel in the INGENYphorU is as easy as gel casting will ever be. The gel sandwich is placed in the electrophoresis cassette during gel casting; the U-shaped spacer closes the gel sandwich on 3 sides (left, right and bottom). The gel can then be cast with the comb in place. After polymerization, just press down the spacer to open the bottom of the gel to the electrical field, load your samples and run the gel. The INGENYphorU-2x2 holds 2 28 x 18 cm gels, allowing the analysis of 96 samples in one run. The 48-well combs are 8-channel pipette compatible. The 18 cm long gel allows you to analyze several different fragments in one lane, giving you even more throughput. The cassette is also available as INGENYphorU-2 with single gel capacity. All cassettes come with glass plates, combs (20-well, 32-well, 38-well or 48-well), U-shaped spacers and a dummy glass plate (INGENYphorU-2x2 only). The perpendicular adapter kit lets you run 4 perpendicular gradient gels simultaneously. Terminals are titanium to avoid corrosion, electrodes are platinum iridium alloy for extra strength. The cassette is manufactured in durable acrylic glass. The electrophoresis cassette is used with the INGENYphorU buffer system. Using approximately 17 L of buffer, the temperature is kept constant to within 0.1°C. This is an important feature as temperature is a very critical parameter. The buffer may be used for several runs before it needs to be replaced.
For convenience, the system is equipped with the following features: buffer flow control from the outside, digital display of actual or set temperature, low voltage (12 V) during sample loading, fill marks for buffer levels, transparent walls which allow monitoring of the gel while it runs, a silent self-priming circulation pump, integrated buffer lines and power leads, and a valve for buffer draining.
The system also has all of the necessary safety features. The INGENYphorU buffer system is made of durable acrylic glass and has high-temperature and low-level cut offs to offer protection over the entire temperature range. Electrophoresis voltage is only available with the lid in place; removing the lid will cut the voltage.
This numerous benefits of this system include: 96-well capacity, 100% leak-free gel casting, reproducible gel casting which enhances inter-gel comparison, accurate temperature control in both gels, constant temperature during gel loading, titanium terminals, strong electrodes (platinum-iridium alloy), comes with a reasonable gradient former and longer gels which allow for the separation of several fragments per lane.
But it also has a few annoying quirks, like getting rid of bubbles trapped in the bottom of the gel. Also, it has a very large buffer volume requirement (17 L). Although this probably helps with temperature stability, it is somewhat annoying in terms of buffer preparation.
We mainly use this instrument to perform Denaturing Gradient Gel Electrophoresis (DGGE) experiments for the diagnosis of colorectal cancer, to detect point mutations, microdeletions, or microinsertions found in cancer patients. The principle of this technique is to separate DNA strands based on their actual base composition (i.e. the ratio of GC to AT base pairs that make up a particular segment of DNA). This is accomplished by exposing the DNA to a gradient of denaturant at elevated temperatures within a polyacrylamide gel. As the DNA sample progresses through the gel, from a low denaturant concentration to a higher one, it starts to melt at varying points. The higher the GC content of the sample, the harder it is to melt. Thus, the DNA sample is able to progress further into the gel before stopping. Samples with lower GC content melt more rapidly in comparison. Therefore, they progress more slowly within the gel, thus becoming separated from the other faster moving strands of DNA.
This instrument can also be used for other applications, such as CDGE (Constant Denaturing Gel Electrophoresis), TGGE (Temperature Gradient Gel Electrophoresis), SSCP (Single Stranded Conformation Polymorphism), HA (Heteroduplex Analysis), PTT (Protein Truncation Test).
Tarun Jain
Application Specialist
Biotron Healthcare
Molecular Biology Laboratory