Journal
/
/
Echtzeit-Tracking der DNA-Fragment-Trennung durch Smartphone
Real-time Tracking of DNA Fragment Separation by Smartphone
JoVE Journal
Biochemie
Author Produced
Zum Anzeigen dieser Inhalte ist ein JoVE-Abonnement erforderlich.  Melden Sie sich an oder starten Sie Ihre kostenlose Testversion.
JoVE Journal Biochemie
Real-time Tracking of DNA Fragment Separation by Smartphone

Echtzeit-Tracking der DNA-Fragment-Trennung durch Smartphone

Please note that all translations are automatically generated. Click here for the English version.

14,597 Views

06:58 min

June 01, 2017

DOI:

06:58 min
June 01, 2017

14567 Views

Transkript

Automatically generated

The passage introduces a new system about SNAP gel electrophoresis. This system can not only greatly reduce the chemical conception experiment time, but also can realize real-time tracking in a separation in the agarose gel by smartphone. Those three pictures represent a typical result after gel electrophoresis of 50, 100, and 1, 000 base pairs in a ladder.

Each DNA fragment was well separated. SNAP gel electrophoresis is the most inventive way for DNA fragment separation, and, thus, it is deemed a vessel tutor in biochemical and biological analysis. This time I will show you a new method about SNAP gel electrophoresis experiment.

Okay, let’s begin. First, we weigh out 0.1 gram of agarose gel and put it into a flask, a TPE buffer into the flask. Volume of the prepared solution should be not more than one third of the capacity of the flask.

Seal the flask with preservative fume, and then the agarose buffer mixture was heated by a microwave. Prior to putting the flask into microwave, we make some holes in the preservative fume in case of explosion. Hold the mounted agarose solution into the channels in the SNAP gel electrophoresis tube.

Lay the comb into the solution. Mounted solution will cool down and become gel state at room temperature some minutes later. Take out of the comb to create wells and add TBE Buffer to.

We have constructed a simple real time tracking DNA fragment separation system to observe the motion state of DNA fragments by smartphone. First mix the DNA ladder with SYBR Green One by centrifuge. Load the mixture into the well in the agarose gel.

Second, turn on the LED light source and place a band-pass filter above the LED then put the electrode into the two lanes of the chip. Next, put other band-pass filter above it. Wait on the power source.

Set the electric wattage at 180 watt. Turn on smartphone to record the DNA fragment separation process. For example we use smartphone to track separation of 50 base pairs DNA ladder.

SNAP gel electrophoresis is the most effective way for DNA fragment separation. However, abandoned experiments indicate that SNAP gel electrophoresis was restricted by the full length for problems. One:the separations take many hours and even days.

Two:the chemical perception is very high. Three:as it requires a lot of apparatus. Four:the gel imaging system can only observe the separated DNA fragments when the experiment is finished.

Furthermore, Iridium and Bromine which is commonly used in SNAP gel electrophoresis is mutagenic and cancerogenic. After the experiment, each DNA fragment was well separated. Furthermore we have separated the same samples in the four channels of the SNAP gel electrophoresis chip and the issue that the DNA fragments was the same size moved the same distance in each experiment.

From the picture, DNA fragments was separated well although there were doubting DNA fragments looks white in figure B.It can be improved by optimizing the parameters of the chip and the concentration of the agarose gel. After watching this video, you must see our new message about to SNAP gel electrophoresis. Also the resulting DNA fragments looks white.

This can be improved by optimizing the parameters of your SNAP gel electrophoresis chip and the concentration of the agarose gel. What important point is, that you must be very careful during intakes paramount because it needs high voltage and it can become reagent.

Summary

Automatically generated

Traditionelle Plattengelelektrophorese (SGE) Experimente erfordern eine komplizierte Apparatur und einen hohen chemischen Verbrauch. Diese Arbeit stellt ein Protokoll vor, das eine kostengünstige Methode zur Trennung von DNA-Fragmenten innerhalb eines kurzen Zeitrahmens beschreibt.

Verwandte Videos

Read Article