Blue:RNA-Seq Experiments:08052013

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Purpose[edit]

  • Sort single human HSCs (Zhili Rong) per well for totoRNAseq analysis


Prepare 5x HiMg RT buffer:[edit]

' Stock Final (5x) Vol for 1 ml
Tris-HCl pH8.3 1M 250mM 250ul
KCl 1M 375mM 375ul
MgCl2 1M 60mM 60ul
dH2O 315ul


Prepare RT Sort Buffer:[edit]

Component Volume
5xRT Buffer 500ul
10% Triton-X100 10ul
40U/ul Rnase Inhibitor 25ul
ERCC spike-in RNA (1:10^5) 10ul
Water 455ul
total 1000ul


Plate Layout:[edit]

' 1 2 3 4 5 6 7 8 9 10 11 12
A 10cell(1) 10cell(9) Sc(17) Sc(25) Sc(33) Sc(41) 10cell(49) 10cell(57) Sc(65) Sc(73) Sc(81) Sc(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) Sc(24) Sc(32) UHRR(40) NC(48) Sc(56) Sc(64) Sc(72) Sc(80) UHRR(88) NC(96)


Sc=Single Human HSC (teratoma-derived); (#)=Indx


08-05-2013: One plate per sort condition (CD34+/GFP- and CD34+/GFP+) per tumor (N1_AI and N3_AI_OP9). i.e. 4 plates in total.


08-06-2013: One plate per sort condition* (CD34+/GFP- and CD34+/GFP+) per tumor (N4_HSC_AI, N5_HSC_AIT2, N6_HSC_AIOP9). *one extra plate for N5_HSC_AIT2 GFP+. i.e. 7 plates total.