User:Tim Henry/Notebook/D1S80/2012/02/25

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D1S80 w/ LB buffer (attempt #2)

PCR on 20120224. Tim and Noah products did not appear, other two lanes not clear anyway.

1.2% gel w/ LB buffer

65mL + .78g agarose.

Used smaller gel tray / less lanes, but bigger wells to hold more

280v for ~40mins

Looked pretty bad, ladder not even clearly visible. TAE seemed to perform much better. May be due to salt concentration and not using supplied loading buffer (Orange G). PCR mix has loading already included, ladder does not and could try separate runs with just ladder until clear. Possibly lower voltage regardless of what doc says. Also should be noted that smaller gel tray does not span all of electrophoresis tank, and black electrode does not span full length (1/4 to 1/2) either. May be related.

It is very tempting to switch back to TAE at this point or try TBE, but since I already have the LB should experiment more to go for sharper / readable bands.

Thenry D1S80 20120225.jpg