Alberta/Calender/July
From 2007.igem.org
Schedule Legend: JG: Jason, Mc: Michelle, ED: Erin, NG: Nick G., NK: Nik K., CZ: Celine, AF: Adam, Al: Alex, JB: Jori, JP: Justin, VH: Veronica,
July
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July 1
July 2
July 3
July 4
Lab Notes:
Transformations of: J61003 (vector containing GFP), J23119 (constitutive promotor), E1010 (RFP), J45100 (methyl salicylate), 0034 (RBS). Transform into XL10 gold cells. Plate on Amp.
-ED, JG, ML, MC, VH, AL, CZ
July 5
July 6
Lab Notes:
Transformation of E1010 did not work, it is in a Kan vector. SMRT.
Set up 4X5mL overnights of pSB1A3, J45100, B0034, J61003.
-ED
July 7
Lab Notes:
Glycerol stocks of overnights from July 6. Stored in -80 freezer. Miniprep from overnights started July 6. Stored in "iGEM" freezer box in -20 freezer.
-JP,JB,AL
July 8
Lab Notes:
Transform E1010 and A0500 into XL10 Gold. Plate on Kan.
Pour Kan and Amp plates. Make TBE and TAE buffer.
-ED, JP, MC, VH, CZ
July 9
Lab Notes:
Retransform E1010 and A0050 into XL10Gold cells using new plates that were poured Sunday.
-ED, VH, JG
July 10
Lab Notes:
Transformations of E1010 and A0050 were not successful. Tomorrow we will try with a new cell line and enriched media.
-ED, MC, AF, CZ
July 11
Lab Notes:
Third attempt at transforming the Kan resistant BioBricks. Transformed E1010, A0500, and pSB2K3 into XL10 Gold, XL10 Gold with enriched media (Magnesium & Glucose), and DH5a cells. Plated 100uL and the pellet remaining after taking the 100uL aliquot on Kan plates.
Restriction digest of J61003 with Xba and Spe. Run on 0.8% agarose gel. Cut out 2290 band, will gel extract tomorrow.
-ED, MC, NG, JG, VH, CZ
July 12
General Notes:
Calender up and running.
Meeting tonight at 7:00 in CSC 2-49.
Received Chlorobium tepidum, meeting with Dr. Jeff Fuller (with Capital Health) to discuss the use of their anaerobic chambers!! -Justin
Lab Notes:
Transformations with DH5a cells worked, but we will need to select for single colonies tonight.
Streak for single colonies of E1010, A0500, J61003.
Purification of gel slice of digested J61003.
-ED, JG, AL, JB, MC, CZ
July 13
General Notes:
Everyone who is coming tonight please be present 7:00pm sharp
Lab Notes:
Started 4X5mL overnights of E1010 and I0050 at 8:30 am. Should be ready for miniprep tonight.
-ED, JB
No growth in over nights - left these shaking in 37 incubator because they may be "slow growers"
Started new overnights of E1010, I0500, and psb2k3 (all DH5alpha) at 2200hrs incubating in the incubator by the autoclave - plates were also streaked (&labeled corresponding to the tubes)
Also made overnights for massive colonies seen on 48hr growth of E1010 in XL
Made some Kan plates
-MC, JP, JG, CZ
July 14
Schedule:
AL,JP,NK
General Notes:
If you are scheduled during the day please be in the lab at 1230hrs (PM) Don't forget that the Bio-Sci doors lock at 1900hrs so if you are trying to get in after 1900hrs call 492-2911
Things to be continued today (CZ)
1. Put LB plates in 4 degree fridge
2. Plate 20 microlitre of DH5 alpha on a kan plate as a control
3. Xba1/EcoRI double digest
NOTE: Sequential digestion is needed. Start with 1 microlitre of Xba1 and 2 microlitre of 10x Tango and incubate for an hour. Then add 1 microlitre of EcoRI and 2.5 microlitre of 10X Tango and incubate for an hour.
4. Miniprep on the colonies picked on July 13 at 10pm. They are in the shaker in the back room. Note the colonies picked were also streaked on kan plates in the 37 degree incubator. Erin's colonies are in the shaker in the front room.
If you need anything else call me(Celine). Number on the blackboard. Thank you.�
<b/>Lab Notes:</b>
JP, AL, NK, JB(?)
1. Completed Mini on E10050 (4), I0500 (2), PSB2K3 (2) samples (the two labeled Erin, are those from July 13 8:30am ON's) In DNA box -20
2. Completed sequential double restriction on J61003 promoter region. Used Xba1 and EcoR1 (see protocol). sample in DNA box -20
3. Placed Kan plates at +4
July 15
Schedule:
Jp,JB,AL
General Notes:
Options:
1. complete double restriction on newly mini'd plasmids (arabinose promoter, RFP)
2. gel purify J61003 restriction products (promoter restriction sample)
3. fundraising letters!
Lab Notes:
1. ran a gel containing the digested products of the J61003 (promoter digest), snapped photo and cut out bands for extraction/purification tomorrow
2. glycerol stocks of Chlorobium tepidum
3. finished grant application forms
- JP, AL, NG, MC
July 16
Schedule:
CZ,VH,AF
General Notes:
1. Because we didn't get into lab till late (because of locked doors), did not complete restriction on newly mini'd plasmids. Can be completed today (isolate arabinose promoter and RFP)
2. All of the restriction product (for J61003 XbaI/EcoR1) were ran and cut - no sample remaining
Lab Notes:
Completed gel extraction of E,X digested J61003. (in box in -20)
Restriction digests of E1010 (X,S) and I0500 (E,x), ran on 0.8% gel. E1010 expected a band at 680bp, I0500 expected a band at 1200 bp. Got two positive lanes for E1010: XL1 and XL2 minipreps. Cut out bands and put in -20. No positive for I0500.
-VH, ED, CZ
Quotes of the Day:
"If I had a remote, I'd change the cd."
Anonymous, in response to the skipping music on the cd player, but much to lazy to get up and change it
"I would never drive this tired. But I WILL operate chemicals."
Anonymous
July 17
Schedule:
ED,VH,AF
Lab Notes:
Gel extracted E1010 from yesterday's slices.
Mini-prep on A0500 from a culture labelled with red marker. If anyone knows what cell line is in this culture, please post.
It is I0500 and it is DH5alpha unless otherwise labelled . .. only the E1010 had both DH5alpha and XL10gold overnights -MC
Let's not do anything more with it then, because the DH5as aren't working due to them already having their own Kan resistance. - ED
Ligation of E1010 (RFP) into J61003. Used 4 different ligation conditions. These are sitting on the instructor's bench at the front of the room. These need to be transformed into XL10 Gold and plated on Amp tomorrow. Try transforming 1uL and 10uL.
-ED,JG,VH,AF
Quote of the Day
"Cancer is an urban myth."
July 18
Schedule:
CZ,MC,NG
July 19
Meeting at 7:00, CAB 373.
Schedule:
ED,MC,JG
July 20
Schedule:
JG,ED,MC
July 21
Schedule:
JP,NK,NG
July 22
Schedule:
JP,AL,NK
AL: Sorry guys, but I won't be available this Sunday for the lab due to staff meeting at work, can anyone swap with me?
July 23
July 24
July 25
July 26
Meeting at 7:00, Room TBA.
July 27
July 28
July 29
July 30
July 31
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