"
Team:Edinburgh/Notebook/BRIDGE
From 2010.igem.org
(→BRIDGE) |
|||
| Line 109: | Line 109: | ||
== BRIDGE == | == BRIDGE == | ||
| + | <br><br><br><br> | ||
| + | <b>25/10/10</b> | ||
<br> | <br> | ||
| + | Repetitive failure of purifications with both traditional method and kits, unsure what is going wrong. Have decided to tell Chris. Off to Firbush on Wednesday.<br><br> | ||
| + | <b>01/10/10</b> | ||
| + | <br> | ||
| + | In my absence (at Firbush with the rest of the Biotechnologists) Chris has managed to construct the tnaA UP-cat-sacB-tnaA DOWN segment that we need for the first part of the BRIDGE protocol. I will continue to build the tnaA UP-RFP-tnaA DOWN segment. | ||
| + | <br> | ||
| + | Set up ligations of RFP-DOWN and UP-RFP, left in 16C incubator overnight. | ||
| + | <br><br> | ||
| + | <b>02/10/10</b> | ||
| + | <br> | ||
| + | Lab on a Saturday again, fun fun fun... | ||
| + | <br> | ||
| + | Retrieved and amplified ligations and ran on gel. UP-RFP appears to be significantly shorter than RFP-DOWN and indeed shorter than it ought to be (should be about 2kb, is only around 1.5kb). <br> | ||
| + | Purified both products, then digested RFP-DOWN with XbaI. (Used 2hour incubation time to write presentation for Wednesday).<br> | ||
| + | Set up ligation of UP with RFP-DOWN and left in incubator overnight.<br> | ||
| + | <br> | ||
| + | <b>03/10/10</b> | ||
| + | <br> | ||
| + | Bus to Darwin for 11:20am; enter Darwin; lift to 8th floor; retrieve ligations from waterbath in cold room; walk down to 7th floor; put ligation in freezer; lift to ground floor; leave Darwin; catch 11:40am bus back to central. | ||
| + | <br><br> | ||
| + | <b>04/10/10</b> | ||
| + | <br> | ||
| + | PCR of ligation of UP-RFP-DOWN. Made and ran gel along with purified PCRs from Saturday, just in case they're still not working. | ||
Revision as of 11:40, 4 October 2010
BRIDGE
25/10/10
Repetitive failure of purifications with both traditional method and kits, unsure what is going wrong. Have decided to tell Chris. Off to Firbush on Wednesday.
01/10/10
In my absence (at Firbush with the rest of the Biotechnologists) Chris has managed to construct the tnaA UP-cat-sacB-tnaA DOWN segment that we need for the first part of the BRIDGE protocol. I will continue to build the tnaA UP-RFP-tnaA DOWN segment.
Set up ligations of RFP-DOWN and UP-RFP, left in 16C incubator overnight.
02/10/10
Lab on a Saturday again, fun fun fun...
Retrieved and amplified ligations and ran on gel. UP-RFP appears to be significantly shorter than RFP-DOWN and indeed shorter than it ought to be (should be about 2kb, is only around 1.5kb).
Purified both products, then digested RFP-DOWN with XbaI. (Used 2hour incubation time to write presentation for Wednesday).
Set up ligation of UP with RFP-DOWN and left in incubator overnight.
03/10/10
Bus to Darwin for 11:20am; enter Darwin; lift to 8th floor; retrieve ligations from waterbath in cold room; walk down to 7th floor; put ligation in freezer; lift to ground floor; leave Darwin; catch 11:40am bus back to central.
04/10/10
PCR of ligation of UP-RFP-DOWN. Made and ran gel along with purified PCRs from Saturday, just in case they're still not working.
