Blue:RNA-Seq Experiments:09302013: Difference between revisions

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::'''Note: using 2ul per well for 4 plates
::'''Note: using 2ul per well for 4 plates (two E12.5 and two E10.5)
 


== Universal Sort Buffer: ==
== Universal Sort Buffer: ==

Revision as of 20:51, 30 September 2013

Purpose

  • mNPCs from E12.5 and E10.5 mice were isolated by Yun
  • Sorting of mNPCs (mixed population of cell types) was based on PI negative staining

Smart-Seq Sort Buffer:

Component Stock Final (1x) Volume
Clontech Dilution Buffer 772ul
Rnase Inhibitor 40U/ul 1U/ul 20ul
ERCC spike-in RNA 1:10^5 8ul
total 800ul


Note: using 2ul per well for 4 plates (two E12.5 and two E10.5)

Universal Sort Buffer:

Component Stock Final (1x) Volume
DEPC Water 384.7ul
10% Triton-X100 4ul
Rnase Inhibitor 40U/ul 1U/ul 10ul
ERCC spike-in RNA 1:10^5 5.3ul
total 400ul


Note: using 0.75ul per well for 4 plates and bubbling just prior to sort

Plate Layout:

' 1 2 3 4 5 6 7 8 9 10 11 12
A NC(1) UHRR(9) NC(17) NC(25) UHRR(33) NC(41) NC(49) UHRR(57) NC(65) NC(73) UHRR(81) NC(89)
B Sc(2) Sc(10) Sc(18) Sc(26) Sc(34) Sc(42) Sc(50) Sc(58) Sc(66) Sc(74) Sc(82) Sc(90)
C Sc(3) Sc(11) Sc(19) Sc(27) Sc(35) Sc(43) Sc(51) Sc(59) Sc(67) Sc(75) Sc(83) Sc(91)
D Sc(4) Sc(12) Sc(20) Sc(28) Sc(36) Sc(44) Sc(52) Sc(60) Sc(68) Sc(76) Sc(84) Sc(92)
E Sc(5) Sc(13) Sc(21) Sc(29) Sc(37) Sc(45) Sc(53) Sc(61) Sc(69) Sc(77) Sc(85) Sc(93)
F Sc(6) Sc(14) Sc(22) Sc(30) Sc(38) Sc(46) Sc(54) Sc(62) Sc(70) Sc(78) Sc(86) Sc(94)
G Sc(7) Sc(15) Sc(23) Sc(31) Sc(39) Sc(47) Sc(55) Sc(63) Sc(71) Sc(79) Sc(87) Sc(95)
H Sc(8) Sc(16) 10cell(24) Sc(32) Sc(40) 10cell(48) Sc(56) Sc(64) 10cell(72) Sc(80) Sc(88) 10cell(96)


Sc=Single cell; (#)=Indx


Notes:

  • Sort was done on the FACS ARIA II