Following yesterday's very strange results with the displacement strands, we tested the binding ability of the displacement oligos. Oligos were run on a 4-20% PA gel in TBE for 30 minutes and stained with SYBR gold.
|1||25 bp ladder|
|2||20 pmol T20|
|3||20 pmol S20|
|4||20 pmol A20|
|5||20 pmol R20.20|
|6||20 pmol R20.35|
|7||20 pmol T20 + R20.20|
|8||20 pmol T20 + R20.35|
|9||20 pmol A20 + R20.20 (30 minute incubation)|
|10||20 pmol A20 + R20.35 (30 minute incubation)|
A20, T20 + R20.20, T20 + R20.35 were heated to 95C for 5 minutes and cooled slowly back to room temp for 20 minutes before use.
Yesterday's samples were washed 3X in 250 uL PBS, blocked in 100 uL odyssey blocking buffer for 1 hour, washed 3X, incubated with 200 uL primary antibodies in odyssey blocking buffer, washed 3X, incubated with 100 uL secondary antibodies for 1 hour, washed, and scanned.