9:45 A – 7:30 P
General
- Lab meeting (notes)
Gel Free Electrophorysis
- commercial setup
- testing ULR Generuler ladder in low weight and high weight cut-off bags.
- Ran separation for 15 min at 80 V
- Testing on 5% gel
- Melted 5% gel running 15 min in Sodium Borax buffer at 300 V.
- Try again at 150V, 50 min.
- Gel looks good. 50 bp band is partially preserved. smaller bands are lost.
- Singled stranded DNA should have half the molecular weight, maybe we’ll be losing our probe if the 50 bp band is getting depleted.
- Also, recovered only 20 uL of 60 uL loaded into bag. And the concentration clearly dropped.
- Some of the volume loss is for setup reasons — some sample wicks up the sides of the closed bag when you pinch it shut. Maybe a bigger internal spacer could prevent this.
- At ~660 Da per dsDNA bp, 50 bp should not make it through the 5,000 da cutoff (33kda). The 35 bp band should be 23 kda. Maybe the charge is too high? Or the solution too hot?