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,

Contents 1 July 2 July 1 3 July 2 4 July 3 5 July 4 6 July 5 7 July 6 8 July 7 9 July 8 10 July 9 11 July 10 12 July 11 13 July 12 14 July 13 15 July 14 16 July 15 17 July 16 18 July 17 19 July 18 20 July 19 21 July 20 22 July 21 23 July 22 24 July 23 25 July 24 26 July 25 27 July 26 28 July 27 29 July 28 30 July 29 31 July 30 32 July 31

July

To August 2007
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July 1

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July 2

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July 3

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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

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July 5

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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

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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

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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

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July 9

Lab Notes:

Retransform E1010 and A0050 into XL10Gold cells using new plates that were poured Sunday.

-ED, VH, JG

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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

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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

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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

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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

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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. SpeI (BcuI)/EcoRI double digest
NOTE: Sequential digestion is needed. Start with 1 microlitre of BcuI 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

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

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July 15

Schedule:

Jp,JB,AL

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July 16

Schedule:

ED,VH,AF

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July 17

Schedule:

CZ,JG,AF

Celine is unable to make it on Tuesday night. Erin, can I trade with you on Monday night?

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July 18

Schedule:

CZ,MC,NG

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July 19

Meeting at 7:00, Room TBA.

Schedule:

ED,MC,VH

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July 20

Schedule:

JG,ED,MC

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July 21

Schedule:

JP,NK,NG

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July 22

Schedule:

JP,AL,NK

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July 23

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July 24

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July 25

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July 26

Meeting at 7:00, Room TBA.

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July 27

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July 28

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July 29

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July 30

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July 31

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