Team:Macquarie Australia/Protocols/GibsonAssembly
From 2012.igem.org
(Difference between revisions)
(→Gibson Assembly Protocol) |
|||
Line 4: | Line 4: | ||
- | Before entering the lab, the assigned Gibson Assembly team for the day organised agar plate requirements, concentration calculations and equipment requirements. | + | <center>Before entering the lab, the assigned Gibson Assembly team for the day organised agar plate requirements, concentration calculations and equipment requirements. <center/> |
- | Each tube was labelled with antibiotic (C = chloramphenicol, K = kanamycin, A = ampicillin). | + | <center>Each tube was labelled with antibiotic (C = chloramphenicol, K = kanamycin, A = ampicillin). |
{| border="3" cellpadding="4" cellspacing="0" align="center" style="width: 100%; height: 400px" | {| border="3" cellpadding="4" cellspacing="0" align="center" style="width: 100%; height: 400px" |
Revision as of 10:39, 5 September 2012
Gibson Assembly Protocol
<center>Each tube was labelled with antibiotic (C = chloramphenicol, K = kanamycin, A = ampicillin).
#1C | #2K | #2A | #3K | #3A | #4C | #5K | #5A | |
---|---|---|---|---|---|---|---|---|
1. Addition of 20ng Gene Block Fragments in appropriate tube | 2µl Hemo_T7A | 2µl Hemo_A | 2µl Hemo_A | 2µl Deino_A | 2µl Deino_A | 2µl Agro_T7A | 2µl Agro_A | 2µl Agro_A |
2µl Hemo_B | 2µl Hemo_B | 2µl Hemo_B | 2µl Deino_B | 2µl Deino_B | 2µl Agro_B | 2µl Agro_B | 2µl Agro_B
| |
nil | nil | nil | 2µl Deino_C | 2µl Deino_C | 2µl Agro_C | 2µl Agro_C | 2µl Agro_C
| |
nil | nil | nil | 2µl Deino_D | 2µl Deino_D | 2µl Agro_D | 2µl Agro_D | 2µl Agro_D
| |
nil | nil | nil | 2µl Deino_E | 2µl Deino_E | 2µl Agro_E | 2µl Agro_E | 2µl Agro_E | |
2. Addition of 0.05 pmol of vector | 2.7µl PSB-1C3 | 2.9µl PSB-1K3 | 2.8µl PSB-1A3 | 2.9 µl PSB-1K3 | 2.8 µl PSB-1A3 | 2.7µl PSB-1C3 | 2.9µl PSB-1K3 | 2.8µl PSB-1A3 |
3. Addition of Gibson Master Mix (µl) | 10 | 10 | 10 | 12.9 | 12.8 | 12.7 | 12.9 | 12.8 |
4. Addition of deionised H2O | 3.3µl | 3.1µl | 3.2µl | nil | nil | nil | nil | nil |