Difference between revisions of "Team:UCLA/Notebook/Recombinant Expression"
| Line 144: | Line 144: | ||
==Goals== | ==Goals== | ||
| − | + | =Julian's Goals= | |
| + | *Finish Purification of ABD containing construct | ||
| + | *Create hydrogel with ABD and B.mori silk and see if it retains/binds albumin | ||
==Achievements== | ==Achievements== | ||
| − | + | =Julian's Achievements= | |
| + | *Cloned the ABD construct and begun purification methods | ||
==What to accomplish next== | ==What to accomplish next== | ||
| − | + | =Julian= | |
| + | *Test albumin binding properties of the ABD silk construct | ||
| + | *test hydrogel formation and albumin retention | ||
==Raw lab notebook entries== | ==Raw lab notebook entries== | ||
{{#calendar: year=2015 | month= may | title = Team:UCLA/Notebook/Recombinant_Expression | query=preload=Template:UCLA}} | {{#calendar: year=2015 | month= may | title = Team:UCLA/Notebook/Recombinant_Expression | query=preload=Template:UCLA}} | ||
Revision as of 18:02, 29 May 2015
Recombinant Silk Functionalization
mm/dd - mm/dd (most recent weeks on top)
mm/dd: Entry title/short description (most recent days on top)
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mm/dd: Sample Entry
Today we began cloning our GFP.
- PCR'd off template
- Ran gel
- Restriction digest
- Ligated into backbone
For our ligation, we made the following modifications:
- Tried it with newly bought ligase
- Left reaction overnight instead of 2 hrs
- Vector to insert ratio was 1:5 instead of 1:3
PCR Reaction:
| Component | Volume |
| 5X Q5 Reaction Buffer | 5 |
| 10 mM dNTPs | 0.5 |
| 10 uM Forward (primer 3/7) | 1.25 |
| 10 uM Reverse (primer 8) | 1.25 |
| Template (diluted to 1ng/uL) | 0.5 |
| Q5 High Fidelity DNA Polymerase | 0.25 |
| Nuclease Free Water | 16.25 |
Gel: Lot of bands, all at correct sizes
Contents
Goals
Julian's Goals
- Finish Purification of ABD containing construct
- Create hydrogel with ABD and B.mori silk and see if it retains/binds albumin
Achievements
Julian's Achievements
- Cloned the ABD construct and begun purification methods
What to accomplish next
Julian
- Test albumin binding properties of the ABD silk construct
- test hydrogel formation and albumin retention
Raw lab notebook entries
| May | ||||||
| M | T | W | T | F | S | S |
| 1 | 2 | 3 | ||||
| 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| 11 | 12 | 13 | 14 | 15 | 16 | 17 |
| 18 | 19 | 20 | 21 | 22 | 23 | 24 |
| 25 | 26 | 27 | 28 | 29 | 30 | 31 |
| June | ||||||
| M | T | W | T | F | S | S |
| 1 | 2 | 3 | 4 | 5 | 6 | 7 |
| 8 | 9 | 10 | 11 | 12 | 13 | 14 |
| 15 | 16 | 17 | 18 | 19 | 20 | 21 |
| 22 | 23 | 24 | 25 | 26 | 27 | 28 |
| 29 | 30 | |||||
| July | ||||||
| M | T | W | T | F | S | S |
| 1 | 2 | 3 | 4 | 5 | ||
| 6 | 7 | 8 | 9 | 10 | 11 | 12 |
| 13 | 14 | 15 | 16 | 17 | 18 | 19 |
| 20 | 21 | 22 | 23 | 24 | 25 | 26 |
| 27 | 28 | 29 | 30 | 31 | ||
| August | ||||||
| M | T | W | T | F | S | S |
| 1 | 2 | |||||
| 3 | 4 | 5 | 6 | 7 | 8 | 9 |
| 10 | 11 | 12 | 13 | 14 | 15 | 16 |
| 17 | 18 | 19 | 20 | 21 | 22 | 23 |
| 24 | 25 | 26 | 27 | 28 | 29 | 30 |
| 31 | ||||||
| September | ||||||
| M | T | W | T | F | S | S |
| 1 | 2 | 3 | 4 | 5 | 6 | |
| 7 | 8 | 9 | 10 | 11 | 12 | 13 |
| 14 | 15 | 16 | 17 | 18 | 19 | 20 |
| 21 | 22 | 23 | 24 | 25 | 26 | 27 |
| 28 | 29 | 30 | ||||