NYMU Taipei/Lab Notes/2007 9 27
From 2007.igem.org
(Difference between revisions)
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| - | * After re-culture in 20-block plate,randomly pick 6 blocks from CinR+HSL+D-term plate | + | * '''CinR+HSL+D-term check''' |
| - | * PCR check using VF2, VR primers | + | ** After re-culture in 20-block plate,randomly pick 6 blocks from CinR+HSL+D-term plate |
| - | * PCR total 50 uL, 10 uL for GEL separation | + | ** PCR check using VF2, VR primers |
| + | ** PCR total 50 uL, 10 uL for GEL separation | ||
[[Image:NYMU Taipei 20070927 CinR-HSL-D-term plate culture test.jpg]] | [[Image:NYMU Taipei 20070927 CinR-HSL-D-term plate culture test.jpg]] | ||
| - | * sample at each lane | + | ** sample at each lane |
| - | ** 1Kb ladder | + | *** 1Kb ladder |
| - | ** block #1 (insert band appears) | + | *** block #1 (insert band appears) |
| - | ** block #2 (insert band appears, and one extra band around 400 bp) | + | *** block #2 (insert band appears, and one extra band around 400 bp) |
| - | ** block #3 (insert band appears) | + | *** block #3 (insert band appears) |
| - | ** block #4 (insert band appears) | + | *** block #4 (insert band appears) |
| - | ** block #5 (insert band does not appear, only band around 400 bp) | + | *** block #5 (insert band does not appear, only band around 400 bp) |
| - | ** block #6 (insert band does not appear, only band around 400 bp) | + | *** block #6 (insert band does not appear, only band around 400 bp) |
| - | * expect results: one band at 1,623 bp is our insert | + | ** expect results: one band at 1,623 bp is our insert |
| - | ** insert = RBS+CinR (2,843 - 2,053) + RBS+HSL (2,783 - 2,053) + D-term (3,266 - 3,163) | + | *** insert = RBS+CinR (2,843 - 2,053) + RBS+HSL (2,783 - 2,053) + D-term (3,266 - 3,163) |
| - | ** insert = 790 + 730 + 103 = 1,623 | + | *** insert = 790 + 730 + 103 = 1,623 |
| - | * extra band at 400 bp might be plasmid failed to be correctly ligated | + | ** extra band at 400 bp might be plasmid failed to be correctly ligated |
| - | ** PCR product length without any insert = (3,189 - 3,049) + 176 = 316 | + | *** PCR product length without any insert = (3,189 - 3,049) + 176 = 316 |
---- | ---- | ||
* '''References''' | * '''References''' | ||
** [http://partsregistry.org/Image:PsB1AK3_map.jpg vector map of pSB1AK3 and VF2 VR primers] | ** [http://partsregistry.org/Image:PsB1AK3_map.jpg vector map of pSB1AK3 and VF2 VR primers] | ||
Revision as of 09:19, 27 September 2007
- CinR+HSL+D-term check
- After re-culture in 20-block plate,randomly pick 6 blocks from CinR+HSL+D-term plate
- PCR check using VF2, VR primers
- PCR total 50 uL, 10 uL for GEL separation
- sample at each lane
- 1Kb ladder
- block #1 (insert band appears)
- block #2 (insert band appears, and one extra band around 400 bp)
- block #3 (insert band appears)
- block #4 (insert band appears)
- block #5 (insert band does not appear, only band around 400 bp)
- block #6 (insert band does not appear, only band around 400 bp)
- expect results: one band at 1,623 bp is our insert
- insert = RBS+CinR (2,843 - 2,053) + RBS+HSL (2,783 - 2,053) + D-term (3,266 - 3,163)
- insert = 790 + 730 + 103 = 1,623
- extra band at 400 bp might be plasmid failed to be correctly ligated
- PCR product length without any insert = (3,189 - 3,049) + 176 = 316
- sample at each lane
- References
- [http://partsregistry.org/Image:PsB1AK3_map.jpg vector map of pSB1AK3 and VF2 VR primers]
