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Agarose gel

Notes  Comments

agarose gel

agarose, 1x buffer




1. take flask ~1.5-2x volume;
2. weight agarose (in flask);
3. add 1x TAE (or 1x TBE) buffer (with EtBr);
4. weight flask with agarose and buffer, write it on the flask;
5. close flask by paper towel;
6. heat to boiling and 2 min more;
7. weight flask and restore the original weight by H₂O;
8. put flask on shaiker and wait until temperature drops to 50-60°C;
9. pour spacers, wait until agarose became solid (2-5 min);
10. pour agarose into plate, wait until agarose became solid (20-40 min);
11. use plate within 2 hours, or wrap into Saran wrap and store at +4°C (up to 2 months);





Gel volume for different plites

plate volume 11x18cm2 140ml 11x11cm2 110ml 8x12.5cm2 90ml

Notes

• Combs may be removed in advance from normal agarose with >1% concentration. High-resolution agarose, low-concentration gels: combs should be removed just before use under runing buffer.




• Combs may be removed in advance from normal agarose with >1% concentration. High-resolution agarose, low-concentration gels: combs should be removed just before use under runing buffer.




• Low-concentration gels (<0.4%):
  • are fragile, run electrrophoresis at +4°C;
  • to handle these gels, pour them on special support film or on 2-3mm layer of 1.5-% agarose;



• High-resolution gels:
  • should solidify at least 2 hous (preferentially at 4°C);
  • are fragile, it worth to check integrity of wells (load ~3µl of 1x Xylene cyanol loading buffer);