User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/04/07

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Ethanol Post-PCR Clean Up

Procedure

1) For each 10 ul of PCR product, prepare the following mixture in strip-cap tube. 1 ul of 3M sodium acetate (NaOAc) pH 5.2 25 ul chilled absolute ethanol. 2) Transfer the PCR product (20ul) into the tube containing the mixture. 3) Vortex the tube and then store it at -20 for 30-50 min. 4) Spin for 30 min at maximum speed 14.000 rpm. 5) Rremove the supernatant carefully and don't disturb the pellet. 6) Wash the pellet with 70% ethanol. And spin the tube at maximum speed for 5 min. 7) Remove the supernatant and then dry the pellet – air dry. 8) Resuspend the pellet in 10 ul in sterile water.

LCH/HCO Products

07April10 Plate.jpg

Sample ID Nucleic Acid Conc. Unit A260 A280 260/280 260/230 Sample Type
lco/hco 1 29.3 ng/µl 0.586 0.394 1.49 0.88 DNA
lco/hco 9 101.6 ng/µl 2.031 1.215 1.67 2.16 DNA
lco/hco 17 68 ng/µl 1.359 0.876 1.55 1.29 DNA
lco/hco 25 89.8 ng/µl 1.797 1.17 1.54 1.32 DNA
lco/hco 33 86.2 ng/µl 1.724 1.144 1.51 1.28 DNA
lco/hco 41 169.1 ng/µl 3.382 2.087 1.62 1.34 DNA
lco/hco 49 179 ng/µl 3.579 2.207 1.62 1.58 DNA
lco/hco 57 110.9 ng/µl 2.217 1.43 1.55 1.33 DNA
lco/hco 65 145 ng/µl 2.9 1.798 1.61 1.51 DNA
lco/hco 73 129.3 ng/µl 2.586 1.619 1.6 1.45 DNA

Average Concentration = 110 ng/ul

Source Plate 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
Ithaca 1 A2 13
Ithaca 1 A3 14
Ithaca 1 A5 15
Ithaca 1 A6 16
Ithaca 1 B1 17
Ithaca 1 B3 18
Ithaca 1 B5 19
Ithaca 1 C1 20
Ithaca 1 D1 21
Ithaca 1 D3 22
Ithaca 1 D4 23
Ithaca 1 D6 24
Ithaca 2 A1 25
Ithaca 2 A2 26
Ithaca 2 A3 27
Ithaca 2 A4 28
Ithaca 2 A5 29
Pennsylvania (PA) B4 30
Tsukuba 1 A4 31
Tsukuba 1 A6 32
Tsukuba 1 B2 33
Tsukuba 1 B4 34
Tsukuba 1 C1 35
Tsukuba 1 C6 36
Tsukuba 3 D2 37
Tsukuba 3 D3 38
Tsukuba 3 D4 39
Woods Hole 1 A6 40
Woods Hole 1 C2 41
Woods Hole 1 C3 42
Yokohama B5 43
Baltimore A3 44
Baltimore A6 45
E. Windsor D4 46
Harvard A1 47
Ithaca 2 A6 48
Pennsylvania (PA) D4 49
Tsukuba 1 B5 50
Yokohama A2 51
Baltimore C1 52
Ithaca 1 A4 53
Pennsylvania (PA) B6 54
Yokohama B2 55
Yokohama A4 56
Ithaca 1 C4 57
Ithaca 1 D5 58
Baltimore C5 59
E. Windsor A2 60
Pennsylvania (PA) B3 61
Ithaca 1 C3 62
Ithaca 1 C6 63
Harvard A4 64
Ithaca 1 B4 65
Ithaca 1 C5 66
Baltimore B2 67
Baltimore B3 68
Baltimore B6 69
E. Windsor C3 70
Pennsylvania (PA) D6 71
Baltimore B5 72
Ithaca 1 C2 73
Pennsylvania (PA) D5 74
Baltimore A5 75
Pennsylvania (PA) B1 76
Boston A1 A
Boston A4 B
Cape Cod (CC) A1 C
Woods Hole 3 A5 D
Woods Hole 1 B3 E
Woods Hole 1 B5 F
Woods Hole 1 C4 G
Woods Hole 2 A6 H
Woods Hole 3 C2 I
Cape Cod (CC) A3 CCA3

Alien Gene LongPCR Ethanol Cleaned

Ethanol Clean # MIX Contents ' Size Gene CCA3 Result Ethanol Cleaned ng/ul Sample Type
1 23 AV10004 FWD AV10007 RVS 8691 AV212 A good 04Mar10 495.3 DNA
2 1 AV10007 FWD AV10011 RVS 9262 AV212 A good 04Mar10 437.4 DNA
3 2 AV10011 FWD AV10016 RVS 11924 AV212 A good 02Mar10 314.9 DNA
4 24 AV10016 FWD AV10020 RVS 7020 AV212 A good 04Mar10 408.3 DNA
5 25 AV10020 FWD AV10025 RVS 11202 AV212 A good 04Mar10 403.1 DNA
6 4 AV10025 FWD AV10028 RVS 6843 AV212 A good 04Mar10 330.8 DNA
7 34 AV10029 FWD AV10034 RVS 12093 AV240 B good 08Mar10 380.5 DNA
8 40 AV10034 FWD AV10039 RVS 14180 AV240 B good 18Mar10 1128.6 DNA
9 50 AV10039 FWD AV10041 FWD 19555 AV240 B good 18Mar10 1888.1 DNA
10 30 EU643474 FWD EU643474 RVS 10062 AV240 B good 04Mar10 206 DNA
11 31 AV10045 FWD AV10047 RVS 6762 AV240 B good 04Mar10 393.3 DNA
12 32 AV10047 FWD AV10049 RVS 12719 AV240 B good 04Mar10 423.1 DNA
13 33 AV10049 FWD AV10053 RVS 9353 AV240 B good 04Mar10 364.3 DNA
14 78 AV10091 FWD AV10092.2 RVS 8127 AV TEL L A good 19Mar10 515.5 DNA
15 85 AV10092.2 FWD AV10094 RVS 7036 AV TEL L A good 01Apr 1353.9 DNA
16 16 AV10094 FWD AV10098 RVS 8600 AV TEL L A good 02Mar10 334.4 DNA
17 134 AV10098 FWD AV10104 FWD 8000 AV TEL L A&B good 01Apr 466.4 DNA
18 51 AV10104 RVS AV10106 RVS 9618 AV_TEL_L_B good 16Mar 430.3 DNA
19 111 AV10106 FWD AV10108.2 RVS 13400 AV_TEL_L_B good 29Mar 218.4 DNA
20 112 AV10107.1 FWD AV10108 RVS 7900 AV TEL L B good 29Mar 214.1 DNA
21 55 AV10108 FWD AV10111 RVS 14846 AV_TEL_L_B good 16Mar 210.7 DNA
22 117 AV10111 FWD AV10113 RVS 8400 AV TEL L B good 29Mar 355.8 DNA
23 128 AV10113 FWD TEL_L_B_3 RVS 11500 AV TEL L B good 01Apr 422.7 DNA
24 131 AV10114 FWD TEL_L_B_4 RVS 8500 AV TEL L B good 01Apr 374.2 DNA