User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/04/27

Alien genes

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Cycle Sequence AMPure Cleaned PCR Product HCO primer only CS Reaction mixture conc. 1x 90 ' Big Dye Terminator 0.5 45 Primer (FWD or RVS) 5uM 0.4 36 DMSO 0.1 9 5X Seq Buffer 5X 0.4 36 Template * ? 4.6 126 1.4ul / RXN * PEG Cleaned PCR product Cycle Seq Program 96C 10 sec 50C 5 sec 60C 4 min x60 CS Plate Source Plate Source Plate Well EX ID CS Plate Source Plate Source Plate Well EX ID A1 Baltimore A1 1 D6 Baltimore A3 44 B1 Baltimore B1 2 E6 Baltimore A6 45 C1 Baltimore C2 3 F6 E. Windsor D4 46 D1 Baltimore C3 4 G6 Harvard A1 47 E1 E. Windsor A1 5 H6 Ithaca 2 A6 48 F1 E. Windsor A3 6 A7 Pennsylvania (PA) D4 49 G1 E. Windsor B1 7 B7 Tsukuba 1 B5 50 H1 E. Windsor B2 8 C7 Yokohama A2 51 A2 E. Windsor C2 9 D7 Baltimore C1 52 B2 Harvard A2 10 E7 Ithaca 1 A4 53 C2 Harvard A3 11 F7 Pennsylvania (PA) B6 54 D2 Ithaca 1 A1 12 G7 Yokohama B2 55 E2 Ithaca 1 A2 13 H7 Yokohama A4 56 F2 Ithaca 1 A3 14 A8 Ithaca 1 C4 57 G2 Ithaca 1 A5 15 B8 Ithaca 1 D5 58 H2 Ithaca 1 A6 16 C8 Baltimore C5 59 A3 Ithaca 1 B1 17 D8 E. Windsor A2 60 B3 Ithaca 1 B3 18 E8 Pennsylvania (PA) B3 61 C3 Ithaca 1 B5 19 F8 Ithaca 1 C3 62 D3 Ithaca 1 C1 20 G8 Ithaca 1 C6 63 E3 Ithaca 1 D1 21 H8 Harvard A4 64 F3 Ithaca 1 D3 22 A9 Ithaca 1 B4 65 G3 Ithaca 1 D4 23 B9 Ithaca 1 C5 66 H3 Ithaca 1 D6 24 C9 Baltimore B2 67 A4 Ithaca 2 A1 25 D9 Baltimore B3 68 B4 Ithaca 2 A2 26 E9 Baltimore B6 69 C4 Ithaca 2 A3 27 F9 E. Windsor C3 70 D4 Ithaca 2 A4 28 G9 Pennsylvania (PA) D6 71 E4 Ithaca 2 A5 29 H9 Baltimore B5 72 F4 Pennsylvania (PA) B4 30 A10 Ithaca 1 C2 73 G4 Tsukuba 1 A4 31 B10 Pennsylvania (PA) D5 74 H4 Tsukuba 1 A6 32 C10 Woods Hole 1 B3 E A5 Tsukuba 1 B2 33 D10 Woods Hole 1 B5 F B5 Tsukuba 1 B4 34 E10 Woods Hole 1 C4 G C5 Tsukuba 1 C1 35 F10 Woods Hole 2 A6 H D5 Tsukuba 1 C6 36 G10 Cape Cod (CC) A3 CCA3 E5 Tsukuba 3 D2 37 H10 Boston A1 A F5 Tsukuba 3 D3 38 A11 Boston A4 B G5 Tsukuba 3 D4 39 B11 Cape Cod (CC) A1 C H5 Woods Hole 1 A6 40 C11 Woods Hole 3 A5 D A6 Woods Hole 1 C2 41 D11 Woods Hole 3 C2 I B6 Woods Hole 1 C3 42 E11 Baltimore A5 75 C6 Yokohama B5 43 F11 Pennsylvania (PA) B1 76 Nuclear Gene Rotifer Culture 454 Amplification Adenita Culture 1-12 EX Takara RXN Mixture conc 1X 6.5 ' Milli Q water 5.15 33.475 Takara EX Buffer 10X 1 6.5 dNTP 2.5uM 0.8 5.2 Primer Mix 20uM 1 EX Taq 5U 0.05 0.325 Template Instagene 2 13 58.5 9.0ul / RXN A PCR cycle 98C 10 sec 60C 1 min x35 Plate 1 Source Plate Well EX ID Baltimore A1 1 Baltimore B1 2 Baltimore C2 3 Baltimore C3 4 E. Windsor A1 5 E. Windsor A3 6 E. Windsor B1 7 E. Windsor B2 8 E. Windsor C2 9 Harvard A2 10 Harvard A3 11 Ithaca 1 A1 12