User:Tara K. Luckau/Notebook/Team ConGen/2010/10/18

Tara's Lab Notebook

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PCR Thermalcycler Testing 2 Mats w/ Tape Description 10µL in each well of full-skirt PCR plate, PCR seal, 2 silicone mats, lab tape on opposite sides Data: 20102018_TKL_CyclerTest2MatTape.tad Results H6, H7 evaporated, but I think it's workable Plan to do first optimization PCR tomorrow! Primer Rehydration, Scun2 Purpose Rehydrate Scun2 primers for testing tomorrow Scun2-F Scun2-R Scun2-F-M13F(-21) Scun2-R-pigtail Calculations Scun2-F 28.3nmol + 141.5 Low TE = 200µM STK Scun2-R 29.2nmol + 146µL Low TE = 200µM STK Scun2-F-M13F(-21) 26.6nmol + 133µL Low TE = 200µM STK Scun2-R-pigtail 30.5nmol + 152.5µL Low TE = 200µM STK stored overnight in fridge to rehydrate completely before making dilution to 20µM dNTP Mix Purpose Mix dNTPs and aliquot for PCR Calculations Have: Invitrogen 100mM dNTP Set (catalog# 10297-018) (250µL of 25µmol in H2O @ 100mM each) Want: 10mM dNTP mix (2.5mM each), 5 aliquots of 1000µL (C1) (V1) = (C2) (V2) (2.5mM) (5000µL) = (100mM) (V2) V2 = 125µL of each dNTP Wet Work 125µL dATP + 125µL dTTP + 125µL dCTP + 125µL dGTP + 4500µL H2O = 5000µL dNTP Mix @ 10mM (2.5mM each dNTP) Used NanoPure H2O from room 325 Aliquots: 1000µL into each of 5 snap-caps; 4 stored in freezer, 1 stored in fridge for immediate use