"
Template:Kyoto/Notebook/Aug 18
From 2013.igem.org
(Difference between revisions)
(→Electrophoresis) |
(→Liquid Culture) |
||
| (9 intermediate revisions not shown) | |||
| Line 29: | Line 29: | ||
|- | |- | ||
|12||8/17 I13504(RBS-GFP-DT) -1||-- | |12||8/17 I13504(RBS-GFP-DT) -1||-- | ||
| + | |} | ||
| + | [[File:Igku Aug18 Gel Extraction before.jpg]]<br> | ||
| + | [[File:Igku Aug18 Gel Extraction1-2.jpg]]<br> | ||
| + | |||
| + | {| class="wikitable" | ||
| + | !Lane||DNA||Enzyme | ||
|- | |- | ||
|13||1kb ladder||-- | |13||1kb ladder||-- | ||
| Line 42: | Line 48: | ||
|18||8/17 I732020(RBS-lacZα-DT) -1||-- | |18||8/17 I732020(RBS-lacZα-DT) -1||-- | ||
|} | |} | ||
| - | |||
| - | |||
[[File:Igku Aug18 Gel Extraction2 before.jpg]]<br> | [[File:Igku Aug18 Gel Extraction2 before.jpg]]<br> | ||
| - | [[ | + | [[File:Igku Aug18 Gel Extraction2-2.jpg]]<br> |
| + | <br> | ||
{| class="wikitable" | {| class="wikitable" | ||
!Name||concentration[µg/mL]||260/280||260/230 | !Name||concentration[µg/mL]||260/280||260/230 | ||
| Line 67: | Line 72: | ||
!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2 | !state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2 | ||
|- | |- | ||
| - | |experiment||J23100||9.0||RBS-lacZα-DT||8.6|| | + | |experiment||J23100||9.0||RBS-lacZα-DT||8.6||3.5 |
|- | |- | ||
| - | |experiment||J23100||9.0||RBS-GFP-DT||7.3|| | + | |experiment||J23100||9.0||RBS-GFP-DT||7.3||3.5 |
|- | |- | ||
| - | |experiment||RBS ||1||lysis3||4.6|| | + | |experiment||RBS ||1||lysis3||4.6||2.8 |
|- | |- | ||
| - | |experiment||Plux ||1||GFP||7.3|| | + | |experiment||Plux ||1||GFP||7.3||3.5 |
|- | |- | ||
| - | |experiment||J23100 ||9.0||--||--|| | + | |experiment||J23100 ||9.0||--||--||3.5 |
|- | |- | ||
| - | |experiment||RBS||1.0||--||--|| | + | |experiment||RBS||1.0||--||--||0.5 |
|- | |- | ||
| - | |experiment||Plux||1.0||--||--|| | + | |experiment||Plux||1.0||--||--||0.5 |
|} | |} | ||
incubate 16°C 1hour | incubate 16°C 1hour | ||
| Line 110: | Line 115: | ||
|1||100bp ladder||--||-- | |1||100bp ladder||--||-- | ||
|- | |- | ||
| - | |2||lysis1 || | + | |2||lysis1 ||XbaI||PstI |
|- | |- | ||
| - | |3||lysis1|| | + | |3||lysis1||XbaI||-- |
|- | |- | ||
| - | |4||lysis1||--|| | + | |4||lysis1||--||PstI |
|- | |- | ||
|5||lysis1||--||-- | |5||lysis1||--||-- | ||
|- | |- | ||
| - | |6||lysis2|| | + | |6||lysis2||XbaI||PstI |
|- | |- | ||
| - | |7||lysis2|| | + | |7||lysis2||XbaI||-- |
|- | |- | ||
| - | |8||lysis2||--|| | + | |8||lysis2||--||PstI |
|- | |- | ||
|9||lysis2||--||-- | |9||lysis2||--||-- | ||
| Line 149: | Line 154: | ||
|94°C||94°C||55°C||68°C||-- | |94°C||94°C||55°C||68°C||-- | ||
|- | |- | ||
| - | |5min||30s||30s||1min|| | + | |5min||30s||30s||1min||30 |
|} | |} | ||
</div> | </div> | ||
| Line 162: | Line 167: | ||
|7/8 LB Kana ||Plusgrow(+Kana) | |7/8 LB Kana ||Plusgrow(+Kana) | ||
|- | |- | ||
| - | |8/16 Pbad araC||Plusgrow(+Kana) | + | |8/16 Pbad araC||Plusgrow(+Kana) |
|} | |} | ||
</div> | </div> | ||
| - | ===Medium Plate=== | + | ===LB Medium Plate=== |
<div class="experiment"> | <div class="experiment"> | ||
<span class="author">No name</span> | <span class="author">No name</span> | ||
Latest revision as of 03:43, 28 September 2013
Contents |
Aug 18
Gel Extraction
| Lane | DNA | Enzyme |
|---|---|---|
| 1 | 100kb ladder | -- |
| 2 | 8/11 J23100 -② | -- |
| 3 | 8/11 J23100 -② | -- |
| 4 | -- | -- |
| 5 | 8/17 K117000(lysis1) -1 | -- |
| 6 | 8/17 K117000(lysis1) -1 | -- |
| 7 | -- | -- |
| 8 | 8/17 K748002(lysis3) -1 | -- |
| 9 | 8/17 K748002(lysis3) -1 | -- |
| 10 | -- | -- |
| 11 | 8/17 I13504(RBS-GFP-DT) -1 | -- |
| 12 | 8/17 I13504(RBS-GFP-DT) -1 | -- |
| Lane | DNA | Enzyme |
|---|---|---|
| 13 | 1kb ladder | -- |
| 14 | 8/17 I0462(RBS-luxR-DT) -1 | -- |
| 15 | 8/17 I0462(RBS-luxR-DT) -1 | -- |
| 16 | -- | -- |
| 17 | 8/17 I732020(RBS-lacZα-DT) -1 | -- |
| 18 | 8/17 I732020(RBS-lacZα-DT) -1 | -- |
| Name | concentration[µg/mL] | 260/280 | 260/230 |
|---|---|---|---|
| 8/14 J231000 | 1.7 | 1.61 | 0.04 |
| 8/17 K748002(lysis3) -1 | 5.4 | 2.39 | 0.45 |
| 8/17 I13504(RBS-GFP-DT) -1 | 3.4 | 3.33 | 0.19 |
| 8/17 I0462(RBS-luxR-DT) -1 | 4.4 | 2.07 | 0.37 |
| 8/17 I732020(RBS-lacZα-DT) -1 | 2.9 | 2.17 | 0.29 |
Ligation
| state | Vector | Inserter | Ligation High ver.2 | ||
|---|---|---|---|---|---|
| experiment | J23100 | 9.0 | RBS-lacZα-DT | 8.6 | 3.5 |
| experiment | J23100 | 9.0 | RBS-GFP-DT | 7.3 | 3.5 |
| experiment | RBS | 1 | lysis3 | 4.6 | 2.8 |
| experiment | Plux | 1 | GFP | 7.3 | 3.5 |
| experiment | J23100 | 9.0 | -- | -- | 3.5 |
| experiment | RBS | 1.0 | -- | -- | 0.5 |
| experiment | Plux | 1.0 | -- | -- | 0.5 |
incubate 16°C 1hour
Restriction Enzyme Digestion
| 8/17K112806(lysis2)-1 | XbaI | PstI | Buffer | BSA | MilliQ | total | |
|---|---|---|---|---|---|---|---|
| 2 cuts | 2.0µL | 1.0µL | 1.0µL | 3.0µL | 3.0µL | 3.0µL | 30 µL |
| 1 cuts | 1.0&mic6/8ro;L | 0.2µL | -- | 1.0µL | 1.0µL | 6.8µL | 10 µL |
| 1 cuts | 1.0µL | -- | 0.2µL | 1.0µL | 1.0µL | 6.8µL | 10 µL |
Electrophoresis
| Lane | Sample | Enzyme1 | Enzyme2 |
|---|---|---|---|
| 1 | 100bp ladder | -- | -- |
| 2 | lysis1 | XbaI | PstI |
| 3 | lysis1 | XbaI | -- |
| 4 | lysis1 | -- | PstI |
| 5 | lysis1 | -- | -- |
| 6 | lysis2 | XbaI | PstI |
| 7 | lysis2 | XbaI | -- |
| 8 | lysis2 | -- | PstI |
| 9 | lysis2 | -- | -- |
| 10 | 100bp ladder | -- | -- |
Colony PCR
| Sample | base pair |
|---|---|
| 8/8 LBKana | -- |
| 8/16 Pbad araC | -- |
| NC | -- |
| PreDenature | Denature | Annealing | Extension | cycle |
|---|---|---|---|---|
| 94°C | 94°C | 55°C | 68°C | -- |
| 5min | 30s | 30s | 1min | 30 |
Liquid Culture
| Sample | medium |
|---|---|
| 7/8 LB Kana | Plusgrow(+Kana) |
| 8/16 Pbad araC | Plusgrow(+Kana) |
LB Medium Plate
| volume | 200ml |
|---|---|
| Bacto Trypton | 2g |
| Bacto yeasy extract | 1g |
| NaCl | 1g |
| Agar Powder | 2g |
Transformation
| Name | Sample | Competent Cells | Total | Plate |
|---|---|---|---|---|
| 8/18 J23100+RBS-LacZα-DT | 2µL | 20µL | 22µL | Amp |
| 8/18 J23100+RBS-GFP-DT | 2µL | 20µL | 22µL | Amp |
| 8/18 J23100+RBS-luxR-DT | 2µL | 20µL | 22µL | Amp |
| RBS+lysis3 | 2µL | 20µL | 22µL | Amp |
| Plux+GFP | 2µL | 20µL | 22µL | CP |
| J23100 | 2µL | 20µL | 22µL | Amp |
| RBS | 2µL | 20µL | 22µL | Amp |
| Plux | 2µL | 20µL | 22µL | CP |
Electrophoresis
| Lane | Sample | Enzyme1 | Enzyme2 |
|---|---|---|---|
| 1 | 100bp ladder | -- | -- |
| 2 | 8/16Pbad araC-1 | -- | -- |
| 3 | 8/16Pbad araC-2 | -- | -- |
| 4 | NC | -- | -- |
