0.8% agarose gel concentration with 100V

Hello

base on the concentration table 0.8% is too low,isnt it?

I use it It is fine

you can also try 0.5% agarose gel I did both of them it s OK

Wow! Thats strage really

many thanks

and what about time and intensity?

I use 1 hour with Promega or Invetrogene ladder

Thanks a lot

I use 1.5% and 0.5XTBE . Be sure to adjust for evaporation after microwaving (2.5min on high). Add just enough buffer to cover the meniscus. Voltage 100-120V and less than 50mA. You will see discrete bands 1000-50bp.

thank you

but when i adjust it on 50mA, the voltage stayed at 45V so i increased it to 100mA but the voltage decreased again and it stayed at 85V. what is the problem in your opinion?

The current is proportional to the area of the gel (including the height of the buffer above the gel) that is perpendicular to the electric field.  The values I suggested are for a 10cm wide by 4cm high (including buffer) gel.  If you have a large gel box, you will have a higher current which is fine as long as the gel does not get above room temperature.

thanks a lot

my gel dimention is the same but i dont know why it doesnt work. now i using 85V, 300A for around 45 min and my bands are acceptable somehow.

If your are making your own TAE buffer, be sure to filter sterilize rather than autoclave.

Hi

Better you can use TBE buffer with 1x conc. and also make 3% agarose gel and calculate how many gram of agarose taken for making gel for 3% agarose based on capacity of your gel casting tray. Voltage set 90 - 105. Try this condition.

thank you. unfortunately the power supply was broken and now its intensity doesnt increase more that 50A. i have used 1x TBE and 2% gel. it was not good enough. nowim using 0.5x TBE and 1.5% agaros gel. voltage 100 And 50A. the bands are not so bad! i used to use autoclave but i will try filter strtilize

thanks all of you