Team:BYU Provo/Team Thermosensor/Week17
From 2011.igem.org
Cheddar3210 (Talk | contribs) (Created page with "{{Template:BYU_Provo}} <html><script> var ele = document.getElementById('liTherm17'); ele.style.fontWeight = "bold"; ele.style.backgroundColor = "#059469"; jQuery(documen...") |
Cheddar3210 (Talk | contribs) |
||
Line 11: | Line 11: | ||
<div id='pageContents'> | <div id='pageContents'> | ||
+ | |||
+ | ==Week 17 (Aug. 7 - 13)== | ||
+ | |||
+ | ===8/8/11=== | ||
+ | |||
+ | Redid overnights to freeze down pBADLAT and pBADLATlacZ | ||
+ | |||
+ | Moved Dr. Grose’s 4 slice plate to the fridge from the 37o to the fridge | ||
+ | |||
+ | ===8/9/11=== | ||
+ | |||
+ | IGM 29- T26-1 (col. 2) (pIG 12) | ||
+ | |||
+ | IGM 30- T26-1 (col. 5) (pIG 13) | ||
+ | |||
+ | IGM 31- T36-2 (col. 7) (pIG 15) | ||
+ | |||
+ | IGM 32- T36-2 (col. 3) ( pIG 16) | ||
+ | |||
+ | Boiled Template (T47-1) placed in freezer | ||
+ | |||
+ | Streaked plates T47-1,2, & 3 from Dr. Grose’s 4 colony plate | ||
+ | |||
+ | T47-1 placed in 37oC at 3:25 pm | ||
+ | |||
+ | T47-2&3 placed in 30oC at 3:25pm (2 plates because T47-2 might have gotten messed up) | ||
+ | |||
+ | These plates are to see if there is any difference in expression of lacZ at 30oC vs. 37o | ||
+ | |||
+ | --Mark | ||
+ | |||
+ | ===8/10/11=== | ||
+ | |||
+ | pBAD PCR, according to standard phusion protocol. Used primers IG37 and IG38 | ||
+ | |||
+ | Looked at 37oC plate compared to 30oC plate. Thermosensor A is white on 30oC plate, but a very dark blue on the 37oC plate. Thermosensors B,C, and D show no apparant difference | ||
+ | |||
+ | Not sure why these thermosensors are acting differently. | ||
+ | |||
+ | Restreaked thermosensor A, and picked several more colonies to compare. Chose colonies and labeled them E-N. Double plated each of these colony streaks one in each 30oC and 37oC, Colony A was repeated as a control to the additional new colonies. | ||
+ | |||
+ | -Julie | ||
+ | |||
+ | ===8/11/11=== | ||
+ | |||
+ | Colonies A, I, and J white at 30oC, and blue at 37oC | ||
+ | |||
+ | Set up overnights of these colonies in 5ml LB-Amp. Incubating at 37oC in shaker overnight | ||
+ | |||
+ | -Addision | ||
+ | |||
+ | ===8/12/11=== | ||
+ | |||
+ | Plasmid preps of A, I, and J got messed up. Re-did with 5ml LB-Amp and put in 37oC shaker. | ||
+ | |||
+ | -Addison | ||
+ | |||
+ | Set up transformation with pIG16 (plasmid with everything but thermosensor) and DH5alpa cells. Doing this to make sure there is no leaky lacZ expression. Std. transformation protocol. (T49-1) | ||
+ | |||
+ | --Julie | ||
+ | |||
+ | ===8/13/11=== | ||
+ | |||
+ | Placed T49-1 and overnights A, I, and J in fridge | ||
+ | |||
+ | --Addison | ||
</div> | </div> |
Latest revision as of 08:49, 24 September 2011
|
Contents |
Week 17 (Aug. 7 - 13)
8/8/11
Redid overnights to freeze down pBADLAT and pBADLATlacZ
Moved Dr. Grose’s 4 slice plate to the fridge from the 37o to the fridge
8/9/11
IGM 29- T26-1 (col. 2) (pIG 12)
IGM 30- T26-1 (col. 5) (pIG 13)
IGM 31- T36-2 (col. 7) (pIG 15)
IGM 32- T36-2 (col. 3) ( pIG 16)
Boiled Template (T47-1) placed in freezer
Streaked plates T47-1,2, & 3 from Dr. Grose’s 4 colony plate
T47-1 placed in 37oC at 3:25 pm
T47-2&3 placed in 30oC at 3:25pm (2 plates because T47-2 might have gotten messed up)
These plates are to see if there is any difference in expression of lacZ at 30oC vs. 37o
--Mark
8/10/11
pBAD PCR, according to standard phusion protocol. Used primers IG37 and IG38
Looked at 37oC plate compared to 30oC plate. Thermosensor A is white on 30oC plate, but a very dark blue on the 37oC plate. Thermosensors B,C, and D show no apparant difference
Not sure why these thermosensors are acting differently.
Restreaked thermosensor A, and picked several more colonies to compare. Chose colonies and labeled them E-N. Double plated each of these colony streaks one in each 30oC and 37oC, Colony A was repeated as a control to the additional new colonies.
-Julie
8/11/11
Colonies A, I, and J white at 30oC, and blue at 37oC
Set up overnights of these colonies in 5ml LB-Amp. Incubating at 37oC in shaker overnight
-Addision
8/12/11
Plasmid preps of A, I, and J got messed up. Re-did with 5ml LB-Amp and put in 37oC shaker.
-Addison
Set up transformation with pIG16 (plasmid with everything but thermosensor) and DH5alpa cells. Doing this to make sure there is no leaky lacZ expression. Std. transformation protocol. (T49-1)
--Julie
8/13/11
Placed T49-1 and overnights A, I, and J in fridge
--Addison