IGEM:Harvard/2006/Adaptamers/Notebook/2006-8-27

8/27

Displacement oligos

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.

Lane	Condition
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.

ELISA

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.