'Blue loading buffer' - (Oct/20/2005 )
i would like to know the important of 'blue loading buffer' and the content of this blue loading buffer to the experiment of electroforesis agarose gel.....someone please answer me....thanks a lot....
blue so you can see
glycerol ~ 30% so it go inside the well
xylene cyanol and bromophenol blue, two components of the loading dye that allows to estimate relative migration. For ex : in a 1%agarose, BPB moves as 300pb and XC moves as 2kpb.
I have a question! How to calculate the size BPB and XC move as in different concentration agarose? For ex: What's the size BPB and XC move as in a 0.8% agarose?
i use bromofenol blue and sukrose for 0,8 % agarose.
i get that recipe from sambrock
i get in the maniatis rates, but here you can find additional information
Here you are :
Agarose %...... XC.............BBP
0.5–1.5......... 4–5 Kb ...400–500 bp
2.0–3.0......... 750 bp ......100 bp
4.0–5.0.........125 bp........ 25 bp
you'll get relative migration in acylamide gels here
Our lab switched several years ago to an Orange G-based loading buffer/tracking dye (at 6X: Orange G (0.25% w/v) in ficoll (15% w/v), glycerol (30% w/v), or sucrose (40% w/v) in 1X electrophoresis buffer).
The Orange G band runs at ~50 bp, and does not show up on photos taken under UV light (like bands of xylene cyanol/bromophenol blue do). It's convenient as it stays out of the way, and is especially useful if you do a lot of small DNA electrophoresis (like small PCR products, for example).
We use this loading buffer exclusively now...