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====Actions for next run==== =====To get larger fragment size===== *This is the first priority *Reduce mixing speed, with lower ALS concentration and longer denature time *Jeffβs suggestion : 133mM of ALS after mixing with cell. *Mixer volume/ (Mixer volume β cell capture volume) = 1.12 --> Required concentration of KOH in prepaired ALS tube for 133mM = '''148.96 mM''' (~ 149mM) {| {{table}} | || | ALS, current || | ALS, new || |- |H2O || |69.5 uL || |74.52 uL || |- |1M DTT || |10 uL || |10 uL |- |5M KOH || |8 uL || |2.98 uL |- |8% Tween20 || |12.5 uL || |12.5 uL |} {| {{table}} | || | NS, current || | NS, new || |- |H2O || |0 uL || |54.9 uL || |- |NS stock || |87.5 uL || |32.6 uL |- |8% Tween20 || |12.5 uL || |12.5 uL |} *NS stock : 4mL of 1M HCl + 6mL of 1M TrisHCl =====To reduce sample loss===== *Push the volume in ring mixer with air, and use MDA master mix filled in mixer and run MDA *For the next device design, add inlet between mixer and compartment for extracting MDA product from ring mixer *Total Compartments volume will be almost matched with the volume of ring mixer, as usual. =====To minimize bias===== *Adjust (decrease) concentration of phi29 and primers. =====Changes for next device design===== *Add inlet channels between mixer and compartment, and one more inlet next to ALS for air input *Slightly increase mixer volume --> will check how much volume will be transferred from current ring mixer to compartments using actual ALS with 10min. mixing
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