From 2009.igem.org
(Difference between revisions)
Latest revision as of 20:49, 21 October 2009
7/14 taq virus protein
Mix: | 50μl
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template | 1μl
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forward primer - | 1μl
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reverse primer - | 1μl
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dNTP | 2μl
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10x buffer | 5μl
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taq | 0.25μl
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ddH20 | 39.75 μl
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Protocol:
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1. | 94oC: 60
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2. 30 cycles | 94oC: 15
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55oC: 20
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72oC: 150
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3. | 72oC: 300
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| 2% agarose, 90V, 30min
Descr | Win length | Fail length
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0: marker 1kb+100bp | |
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1: gp120 full length | 2621bp |
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2: gp120 a-b | 2539bp |
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3: H1N1-HA full length | 1742bp |
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4: H5N1-HA full length | 1742bp |
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5: H1N1-NA full length | 1460bp |
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6: H5N1-NA full length | 1391bp |
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7: H5N1-NA a-b | 1283bp |
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8: Positive Control BBa_E0240 | 1114bp (876+238) | 238bp
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9: H5N1-HA b-c | 911 |
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10:H1N1-NA c-d | 606bp |
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11:H1N1-NA d-e | 495bp |
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12:Positive Control pLac | 437bp |
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13:H5N1-HA d-e | 378bp |
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14:H1N1-NA b-c | 315bp |
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15:H5N1-HA a-b | 285bp |
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16:marker 100bp | |
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Mix: | 50μl
|
template | 1μl
|
forward primer - | 1μl
|
reverse primer - | 1μl
|
dNTP | 2μl
|
10x buffer | 5μl
|
taq | 0.25μl
|
ddH20 | 39.75 μl
|
|
Protocol:
|
1. | 94oC: 60
|
2. 30 cycles | 94oC: 15
|
55oC: 20
|
72oC: 30
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3. | 72oC: 300
|
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| 2% agarose, 90V, 30min
Descr | Win length | Fail length
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0:marker 100bp | |
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1:H5N1-HA c-d | 236bp |
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2:gp120 b-c | 114bp |
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3:H5N1-NA b-c | 126bp |
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4:H1N1-NA a-b | 103bp |
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5:negative control | |
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7/20 pfu virus proteins
Mix: | 50μl
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template | 1μl
|
forward primer - | 1μl
|
reverse primer - | 1μl
|
dNTP | 2μl
|
10x buffer | 5μl
|
pfu | 0.5μl
|
ddH20 | 39.5μl
|
|
Protocol:
|
1. | 94oC: 60
|
2. 30 cycles | 94oC: 15
|
55oC: 20
|
72oC: 120
|
3. | 72oC: 300
|
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| 2% agarose, 90V, 30min
Descr | Win length | Fail length
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0: marker 1kb | |
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1-3: gp120 b-c | 114bp |
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4-6: H5N1-HA c-d | 236bp |
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7:marker 1kb | |
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Mix: | 50μl
|
template | 1μl
|
forward primer - | 1μl
|
reverse primer - | 1μl
|
dNTP | 2μl
|
10x buffer | 5μl
|
pfu | 0.5μl
|
ddH20 | 39.5μl
|
|
Protocol:
|
1. | 94oC: 60
|
2. 30 cycles | 94oC: 15
|
55oC: 20
|
72oC: 120
|
3. | 72oC: 300
|
|
|
| 2% agarose, 90V, 30min
Descr | Win length | Fail length
|
0: marker 1kb | |
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1-2: H5N1_HA a-b | 285bp |
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3-4: H5N1-HA d-e | 378bp |
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5: marker 1kb | |
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6: Positive Control BBa_E0240 | 1114bp (876+238) | 238bp
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7: negative control | |
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Mix: | 50μl
|
template | 1μl
|
forward primer - | 1μl
|
reverse primer - | 1μl
|
dNTP | 2μl
|
10x buffer | 5μl
|
pfu | 0.5μl
|
ddH20 | 39.5μl
|
|
Protocol:
|
1. | 94oC: 60
|
2. 30 cycles | 94oC: 15
|
55oC: 20
|
72oC: 150
|
3. | 72oC: 300
|
|
|
| 2% agarose, 90V, 30min
Descr | Win length | Fail length
|
0: marker 1kb | |
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1-2: H5N1-HA b-c | 911bp |
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3-4: gp120 a-b | 2539bp |
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5: marker 1kb | |
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7/20 taq virus proteins
Mix: | 50μl
|
template | 1μl
|
forward primer - | 1μl
|
reverse primer - | 1μl
|
dNTP | 2μl
|
10x buffer | 5μl
|
taq | 0.25μl
|
ddH20 | 39.75 μl
|
|
Protocol:
|
1. | 94oC: 60
|
2. 30 cycles | 94oC: 15
|
55oC: 20
|
72oC: 120
|
3. | 72oC: 300
|
|
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| 2% agarose, 90V, 30min
Descr | Win length | Fail length
|
0: marker 1kb | |
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1: marker 100bp | |
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2: H1N1-HA full length | 1742bp |
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3: H1N1-NA full length | 1460bp |
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4: H5N1-NA full length | 1391bp |
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5: H5N1-NA a-b | 1283bp |
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6: H1N1-NA c-d | 606bp |
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7: H1N1-NA d-e | 495bp |
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8: H1N1-NA b-c | 315bp |
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9: H1N1-NA d-e | 495bp |
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10: H1N1-NA a-b | 103bp |
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11: Positive Control BBa_E0240 | 1114bp (876+238) | 238bp
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12: negative control | |
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13: marker 100bp | |
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14: marker 1kb | |
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7/21 taq H1N1-HA H5N1-NA gp120b-c
Mix: | 50μl
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template | 1μl
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forward primer - | 1μl
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reverse primer - | 1μl
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dNTP | 2μl
|
10x buffer | 5μl
|
taq | 0.25μl
|
ddH20 | 39.75 μl
|
|
Protocol:
|
1. | 94oC: 60
|
2. 30 cycles | 94oC: 15
|
55oC: 20
|
72oC: 120
|
3. | 72oC: 300
|
|
|
| 2% agarose, 90V, 30min
Descr | Win length | Fail length
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0: marker 1kb | |
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1:- | |
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2: H5N1-NA a-b | 1283bp |
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3: H5N1-NA full length | 1391bp |
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4: H1N1-HA full length | 1742bp |
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5: Positive Control BBa_E0240 | 1114bp (876+238) | 238bp
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6: - | |
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7: marker 100bp | |
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Descr | Win length | Fail length
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0: marker 1kb | |
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1: - | |
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2: - | |
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3: H5N1-NA b-c | 126bp |
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4: gp120 b-c | 114bp |
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5: negative control | |
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6: - | |
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7: marker 100bp | |
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7/22 pfu virus protein
Mix: | 50μl
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template | 1μl
|
forward primer - | 1μl
|
reverse primer - | 1μl
|
dNTP | 2μl
|
10x buffer | 5μl
|
pfu | 0.5μl
|
ddH20 | 39.5μl
|
|
Protocol:
|
1. | 94oC: 60
|
2. 30 cycles | 94oC: 15
|
55oC: 20
|
72oC: 120
|
3. | 72oC: 300
|
|
|
| 2% agarose, 90V, 30min
Descr | Win length | Fail length
|
0: marker 1kb | |
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1-2: H1N1_NA a-b | 103bp |
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3-4: gp120 b-c | 114bp |
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5-6: H1N1-NA b-c | 315bp |
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5-6: marker 100bp | |
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Mix: | 50μl
|
template | 1μl
|
forward primer - | 1μl
|
reverse primer - | 1μl
|
dNTP | 2μl
|
10x buffer | 5μl
|
pfu | 0.5μl
|
ddH20 | 39.5μl
|
|
Protocol:
|
1. | 94oC: 60
|
2. 30 cycles | 94oC: 15
|
55oC: 20
|
72oC: 120
|
3. | 72oC: 300
|
|
|
| 2% agarose, 90V, 30min
Descr | Win length | Fail length
|
0: marker 1kb | |
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1-2:H1N1-NA d-e | 495bp |
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3-4:H5N1-HA c-e | 591bp |
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5-6:H1N1-NA c-d | 606bp |
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7: marker 100bp | |
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Descr | Win length | Fail length
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0: marker 1kb | |
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1-2:H5N1-NA a-c | 1175bp |
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3: Positive Control BBa_E0240 | 1114bp (876+238) | 238bp
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4: negative control | |
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5: marker 100bp | |
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7/23 pfu virus protein
Mix: | 50μl
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template | 1μl
|
forward primer - | 1μl
|
reverse primer - | 1μl
|
dNTP | 2μl
|
10x buffer | 5μl
|
pfu | 0.5μl
|
ddH20 | 39.5μl
|
|
Protocol:
|
1. | 94oC: 60
|
2. 30 cycles | 94oC: 15
|
55oC: 20
|
72oC: 120
|
3. | 72oC: 300
|
|
|
| 2% agarose, 90V, 30min
Descr | Win length | Fail length
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0: marker 1kb | |
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1-2:H1N1-HA | 1742bp |
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3-4:H5N1-HA | 1742bp |
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5-6:H5N1-NA a-b | 1283bp |
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7: Positive Control BBa_E0240 | 1114bp (876+238) | 238bp
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Descr | Win length | Fail length
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0: marker 1kb | |
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1-2:H1N1-NA c-e | 1081bp |
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3-4:H1N1_NA a-c | 398bp |
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5-6:H5N1_NA b-c | 126bp |
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7: negative control | |
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