Low Mass Ladder [invitrogen]: 6 blunt-ended fragments from 100bp to 2000 bp Lambda DNA HindIII (lambda digested by HindIII) [neb]: 23130, 94116, ..., 125 bp
Morning: the big human thrombin experiment.
8% gel run at 120V for 1.5 hours.
lane 1: ladder
lane 2: human thrombin
lane 3: human thrombin + T5
lane 4: human thrombin + denatured T5
lane 5: bovine thrombin
lane 6: bovine thrombin + T5
lane 7: bovine thrombin + denatured T5
lane 8: ladder
lane 9: T5
lane 10: denatured T5
lane 11: denatured T5+ bovine
lane 12: denatured T5+ human
I also attempted to see if the DNA components of our aptamers were capable of binding each other. Each lane contains 40 pmol of oligo or complex. Ran a 12% gel for 1 hour at 120V. Incubation was for 15 minutes total. Denaturation: oligos mixed, heated to 90 C for 5 minutes, then rapidly cooled to 4 C for 5 minutes, followed by 5 minutes at room temperature.
Lane 1: ladder
Lane 2: T20
Lane 3: S20
Lane 4: T20 + S20
Lane 5: T20 + S20; denatured during incubation
Lane 6: T35
Lane 7: S35
Lane 8: T35+S35
Lane 9: T35 + S35; denatured during incubation
Lane 10: T50
Lane 11: S50
Lane 12: T50 + S50 (denaturation condition)
The complementary sections appeared to have come together at a pretty high rate, both for denaturing and non-denaturing conditions.