User:Alexander S Mikheyev/Notebook/rotifer alien genes/2010/06/10

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Contents

Nuclear Gene Adenita Culture 454 Amplification

Re-Amp Instagene Cultures 57, C, D, I

EX Takara RXN Mixture conc 1X 5 ' '
Milli Q water4.1520.75
Takara EX Buffer10X15
dNTP2.5uM0.84
Primer Mix20uM1
EX Taq5U0.050.25
TemplateInstagene315
45
9.0ul / RXN add 1.0ul Primer
PCR cycle
98C10 sec
60C1 min
x35
AMP LaneTemplatePrimerAMP LaneTemplatePrimer
1Instagene 57B316Instagene IB3
2Instagene 57C317Instagene IC3
3Instagene 57D318Instagene ID3
4Instagene 57E319Instagene IE3
5Instagene 57F320Instagene IF3
6Instagene CB326Neg ControlB3
7Instagene CC327Neg ControlC3
8Instagene CD328Neg ControlD3
9Instagene CE329Neg ControlE3
10Instagene CF330Neg ControlF3
11Instagene DB331Pos ControlB3
12Instagene DC332Pos ControlC3
13Instagene DD333Pos ControlD3
14Instagene DE334Pos ControlE3
15Instagene DF335Pos ControlF3
*Positive Control = AV 20ng

Load 2.0ul Sample

Image:10June10.jpg

Nuclear Gene Adenita Culture 454 Amplification QIAquick Cleaning

1) Combined products from primer 2-6 for each culture 2) Cleaned with Qiagen QIAquick PCR product purification kit

http://www.qiagen.com/products/dnacleanup/gelpcrsicleanupsystems/qiaquickpcrpurificationkit.aspx#Tabs=t2

see pg 19

Nanodrop results

Sample ID Nucleic Acid Conc. Unit A260 A280 260/280 260/230 Sample Type Factor
175.2ng/µl1.5040.8231.832.15DNA50
260.5ng/µl1.2110.6651.822.17DNA50
390.6ng/µl1.8121.0141.792.2DNA50
497.2ng/µl1.9451.0711.822.18DNA50
552.8ng/µl1.0550.5841.812.14DNA50
682.6ng/µl1.6530.8921.852.07DNA50
775.3ng/µl1.5060.8131.852.08DNA50
856.5ng/µl1.1290.6341.782.15DNA50
987.5ng/µl1.750.9531.842.23DNA50
1065.2ng/µl1.3050.7251.82.09DNA50
1151.5ng/µl1.0290.5741.791.87DNA50
1244.4ng/µl0.8890.4881.822.14DNA50
1351.9ng/µl1.0390.5731.811.93DNA50
1454.9ng/µl1.0980.61.832.19DNA50
1563.8ng/µl1.2770.7031.822.21DNA50
1655.1ng/µl1.1020.5941.862.27DNA50
1764.6ng/µl1.2920.71.852.25DNA50
1868.3ng/µl1.3660.7411.842.19DNA50
1979.1ng/µl1.5830.8671.832.18DNA50
2058.6ng/µl1.1730.6531.82.14DNA50
2145.1ng/µl0.9020.4981.811.84DNA50
2266.2ng/µl1.3230.7341.82.2DNA50
2370ng/µl1.40.7551.862.19DNA50
2445.3ng/µl0.9060.4991.822.16DNA50
2544.3ng/µl0.8870.4871.822.21DNA50
2666.2ng/µl1.3230.7221.832.14DNA50
2749.8ng/µl0.9950.5451.832.16DNA50
2851.7ng/µl1.0340.5671.822.05DNA50
2945.2ng/µl0.9050.5061.791.96DNA50
3076.8ng/µl1.5350.8371.832.11DNA50
3148.4ng/µl0.9680.5441.782.12DNA50
3237.3ng/µl0.7460.4191.781.96DNA50
3328.7ng/µl0.5740.3291.751.72DNA50
3438.5ng/µl0.7710.4281.81.93DNA50
3539.5ng/µl0.7890.4461.771.85DNA50
3644.2ng/µl0.8830.4791.852.13DNA50
3742.6ng/µl0.8530.4711.812.07DNA50
3832.3ng/µl0.6460.3571.811.92DNA50
3949.1ng/µl0.9820.5461.81.99DNA50
4043.7ng/µl0.8750.4831.811.98DNA50
4148.6ng/µl0.9710.5481.772.08DNA50
4246.2ng/µl0.9250.5021.842.15DNA50
4343.5ng/µl0.870.491.781.96DNA50
4446.7ng/µl0.9340.5161.812.12DNA50
4555.1ng/µl1.1030.5911.871.98DNA50
4652.5ng/µl1.050.5841.82.12DNA50
4737.6ng/µl0.7510.421.792.02DNA50
4850.1ng/µl1.0020.5481.832.07DNA50
4959.7ng/µl1.1940.6521.832.11DNA50
5033.8ng/µl0.6760.3851.751.97DNA50
5142.4ng/µl0.8480.4811.761.93DNA50
5266ng/µl1.3210.7291.812.16DNA50
5381.2ng/µl1.6250.8951.812.06DNA50
5442.1ng/µl0.8420.4761.772.05DNA50
5539ng/µl0.780.4581.71.8DNA50
5628.9ng/µl0.5790.3471.671.91DNA50
5751.6ng/µl1.0310.5771.792.12DNA50
5852.3ng/µl1.0450.6021.742.12DNA50
5957.5ng/µl1.150.6391.82.1DNA50
6057.2ng/µl1.1440.6441.782.06DNA50
6149.4ng/µl0.9890.5771.712.03DNA50
6259.8ng/µl1.1960.6741.781.91DNA50
6342.7ng/µl0.8550.4911.741.99DNA50
6445.5ng/µl0.910.5171.761.8DNA50
6543.8ng/µl0.8770.491.791.91DNA50
6637.4ng/µl0.7480.4191.792.07DNA50
6737.9ng/µl0.7570.4371.741.86DNA50
6831ng/µl0.6190.3681.681.8DNA50
6939.4ng/µl0.7890.4451.772.05DNA50
7044.8ng/µl0.8950.5211.721.63DNA50
7137.8ng/µl0.7550.4441.71.86DNA50
7241.8ng/µl0.8360.4811.742.03DNA50
7364.2ng/µl1.2840.721.782.06DNA50
7440.1ng/µl0.8030.4521.781.82DNA50
7533.6ng/µl0.6730.3821.762.06DNA50
7626.5ng/µl0.5310.311.712DNA50
A51.5ng/µl1.0290.5711.82.17DNA50
B51.6ng/µl1.0330.5751.81.86DNA50
C79.3ng/µl1.5870.8891.792.22DNA50
D55.2ng/µl1.1050.6261.772.01DNA50
E49ng/µl0.980.5611.752.05DNA50
F36.7ng/µl0.7350.4381.681.83DNA50
G51.5ng/µl1.0290.591.742.04DNA50
H48.7ng/µl0.9740.561.741.88DNA50
I61.2ng/µl1.2240.6951.762.05DNA50
CCA372.9ng/µl1.4580.8071.812.11DNA50
A. ricciae105.2ng/µl2.1041.1471.842.21DNA50
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