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Arichard:Reagents/celseq primers
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==CEL-Seq RT primer layout== ===Stock layout=== * Barcoded RT primers are in 48 wells of a 92 well plate at 200 uM each, in the following layout: {| {{table}} | align="center" style="background:#f0f0f0;"|'''Barcode number''' | align="center" style="background:#f0f0f0;"|'''1''' | align="center" style="background:#f0f0f0;"|'''2''' | align="center" style="background:#f0f0f0;"|'''3''' | align="center" style="background:#f0f0f0;"|'''4''' | align="center" style="background:#f0f0f0;"|'''5''' | align="center" style="background:#f0f0f0;"|'''6''' | align="center" style="background:#f0f0f0;"|'''7''' | align="center" style="background:#f0f0f0;"|'''8''' | align="center" style="background:#f0f0f0;"|'''9''' | align="center" style="background:#f0f0f0;"|'''10''' | align="center" style="background:#f0f0f0;"|'''11''' | align="center" style="background:#f0f0f0;"|'''12''' |- | A||1||2||3||4||5||6||7||8||9||10||11||12 |- | B||13||14||15||16||17||18||19||20||21||22||23||24 |- | C||25||26||27||28||29||30||31||32||33||34||35||36 |- | D||37||38||39||40||41||42||43||44||45||46||47||48 |- | E||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty |- | F||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty |- | G||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty |- | H||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty||Empty |- | |} * Serial dilutions are necessary to dilute the RT primers to the correct concentration. ===Diluted primer layout=== # Dilute primers to 32 pmol/ul, 3.2 pmol/ul, and 0.32 pmol/ul, depending on input (Note that each primer must be diluted separately for each sample): ## Add 16 ul of 100 pmol/ul primer to 84 ul H2O --> 100 ul 32 pmol/ul primer. Use 0.5 ul of this per 1 ng total RNA. ## Add 10 ul of 32 pmol/ul primer to 90 ul H2O --> 100 ul 3.2 pmol/ul primer. Use 0.5 ul of this per 100 pg total RNA. ## Add 10 ul of 3.2 pmol/ul primer to 90 ul H2O --> 100 ul 0.32 pmol/ul primer. Use 0.5 ul of this per 10 pg total RNA. {| {{table}} | align="center" style="background:#f0f0f0;"|'''100 ul/well''' | align="center" style="background:#f0f0f0;"|'''1''' | align="center" style="background:#f0f0f0;"|'''2''' | align="center" style="background:#f0f0f0;"|'''3''' | align="center" style="background:#f0f0f0;"|'''4''' | align="center" style="background:#f0f0f0;"|'''5''' | align="center" style="background:#f0f0f0;"|'''6''' | align="center" style="background:#f0f0f0;"|'''7''' | align="center" style="background:#f0f0f0;"|'''8''' | align="center" style="background:#f0f0f0;"|'''9''' | align="center" style="background:#f0f0f0;"|'''10''' | align="center" style="background:#f0f0f0;"|'''11''' | align="center" style="background:#f0f0f0;"|'''12''' |- | Conc||32 pmol/ul||3.2 pmol/ul||0.32 pmol/ul||32 pmol/ul||3.2 pmol/ul||0.32 pmol/ul||32 pmol/ul||3.2 pmol/ul||0.32 pmol/ul||32 pmol/ul||3.2 pmol/ul||0.32 pmol/ul |- | A||#1||#1||#1||#2||#2||#2||#3||#3||#3||#4||#4||#4 |- | B||#5||#5||#5||#6||#6||#6||#7||#7||#7||#8||#8||#8 |- | C||#9||#9||#9||#10||#10||#10||#11||#11||#11||#12||#12||#12 |- | D||#13||#13||#13||#14||#14||#14||#15||#15||#15||#16||#16||#16 |- | E||#17||#17||#17||#18||#18||#18||#19||#19||#19||#20||#20||#20 |- | F||#21||#21||#21||#22||#22||#22||#23||#23||#23||#24||#24||#24 |- | G||#25||#25||#25||#26||#26||#26||#27||#27||#27||#28||#28||#28 |- | H||#29||#29||#29||#30||#30||#30||#31||#31||#31||#32||#32||#32 |- | |}
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