Working on tables, figures.
Yesterday’s Bioanalyzer results only give DNA within selected region. Ran Qubit to get total DNA concentration of sample.
For each sample, used this volume to get 100 ng DNA as input for ligation. Sample T6 not enough DNA for 100ng, so used all available.
Ligation reaction: combined DNA with adapter and T4 ligase
|H2O||to 50 ul|
P1: Rather than make a working stock for each adapter P1, just used 0.5 ul of adapters P1.1 - P1.7 for samples T1 - T7, respectively. Made master mix of universal adapter P2 following ligation molarity calculator
T4 mix: Mixed 0.5 ul T4 ligase with 28 ul buffer. Added 4 ul to each reaction.
Incubated samples for 30 min at room temp, heat kill for 10 min at 65°C, then cooled to 21°C.
Szymańska, Z. & Zylicz, M. (2009). Mathematical modeling of heat shock protein synthesis in response to temperature change. Journal of Theoretical Biology, 259, 562–569.
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