(Difference between revisions)
(September 17)
(September 17)
Line 441: Line 441:
- O/days of Ligations from yesterday<br>
- O/days of Ligations from yesterday<br>
<b>NB: AMP plates are marked with a red streak down the SIDE of the plate. if it does not have this - then it is NOT AMP</b><br>
ML (~10:30-1:00)><br>
To do: miniprep, of DBBEBB and DBBBBE><br>
To do: miniprep, of DBBEBB and DBBBBE><br>
restrict with XBA, PST><br>
restrict with XBA, PST><br>
Line 450: Line 447:
Cut with XBA PST and CUT with SPE PST><br>
Cut with XBA PST and CUT with SPE PST><br>
GEl extract XBA, PST><br>
GEl extract XBA, PST><br>
<b>NB: AMP plates are marked with a red streak down the SIDE of the plate. if it does not have this - then it is NOT AMP</b><br>
ML (~10:30-1:00)><br>

Revision as of 17:57, 18 September 2007



September 2007
Su M Tu W Th F Sa
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

To August 2007
To October 2007
Back to UofA iGEM Home

September 1

Lab Notes
1. 24 Minipreps for Diezel Blaze in B0034 using Fermentas Kit; stored in -20 freezer
2. Lots of colonies from the transformation last night; no colonies on negation control (good) and lots of colonies on positive control (good) and no satellite colonies (also good); no colonies were picked for overnights
3. Run the Buddy+Benny in B00 and Betty+Enny in B00 Pst/Xba gel from last night at 50V for another 30 minutes for better resolution; all the digest worked so I cut out the appropriate bands and stored the gel in -20 freezer for purification later


to the top

September 2

to the top

September 3

to the top

September 4


JP,ML,JG 7:00pm

For the record:
Buddy = BuOH DH 1235 bp
Betty = Bb-bhBu-coa dh 923 bp
Benny = Butyryl coa dh 1184 bp
enny = Enoyl coa Hydratase 860 bp
Diezel Blaze = buAld-Dh 2639 bp

Moving was done this mid-day.
New location M349
JG has key & access
- MC, ED, JG

Evening crew - please do a once over of G308 to double check we have not left anything and to clean up anything extra/ things we should get out of the way. thnx - MC

Night Crew


Set up new lab

Things we need - Water Bath, Scale, disposable 13ml overnight tubes, Tip waste

lab work

Gel purification ran on Betty,enny and boo as well as buddy and benny and is in the -20 in the new lab

To Do list for tomorrow

- Restriction on DB needs to be done - If we want to put it in front of BE or BB then Spe/Pst - If we want to put it behind BE or BB then cut with Xba/Pst - maybe do both (use 4microL DNA for digest)

-complete restriction on BE and BB for SPE and PST (then we can complete ligations BE-BB and BB-BE or we can ligate D.B into BE or BB to make BE-DB or DB-BE or BB-DB or DB-BB- Then we can ligate BE-BB-DB or DB-BE-BB or BE-BB-DB or BB-DB-BE or BB-BE-DB and we are kinda finished!

- If I0500 comes in from calgary it needs to be transformed.

- Transform R0080 which is a back up promoter just in case?

-Note we also have to do sequenceing on BB and BE and DB

Shout out to calgary for saving our necks Shout out to the move in crew to the top

September 5


NG,ED,ML 7:00pm

AM Crew Notes:

finished up cleaning up post-move in
got a 'water bath' - JG calibrated it
prepared BB in Boo & BE in Boo for sequencing - this will be ready for pick up tomorrow
- DB in BOO with PST/XBA
- DB in BOO with PST/SPE
- BB in BOO with PST/SPE
- BE in BOO with PST/SPE
NB: there is a section in the masterbook "figuring stuff out" just a few questions I have from being away so long - don't worry about it we should do a wet lab recap to bring everyone on the same page at our meeting Thrusday - MC
<3 JG & MC

Transformed I0500 from Calgary. Transformed R0080. Ran digestions on gel. Ligated digested DB with BB and BE. (all in Boo)


to the top

September 6


NK ED AL 5:00pm (Unless your crazy enough to show up at 5AM)

To Do:

Goodies will be served during the meeting (7:00pm @ CAB 373), so be sure to show up!

to the top

September 7


AM Crew: MC, JG meeting at 800hrs

Lab Notes:
No growth observed for R0080 overnights; just scrapping it because we have I0500
No growth on transformations of BB+DB and BE+DB ligations therefore . . . religated - see masterbook for more details
O/N with AMP for I0500
Transform religations in to XL10 Gold

NB: JG has key access

To Do:
Mini preps on I0500 overnights

to the top

September 8


Super Awesome Marathon of Productiveness

8-11: JG,NG
Miniprep I0500
DB BE and DB-BB transformations did not work.
Re-ligated DB into BB boo and DB into BE boo

11-2: AL,ML
Re-transform BE+DB and BB+DB

2-5: VH
Gel extraction of I0500, but we missed the step "restricting with Ecori+speI"

5-8: JP,NK
Restrction on I0500 mini's with Ecori and SPeI

to the top

September 9


And Yet another super productive Sunday

8-11: NG and CZ

NG - Nobody Called me about the key last night (and I completely forgot to swing by: was with parents). So I will wait by the lab either , until the next group shows up or someone with a key. Also bioSci is locked but there is an open door by the physcology wing. I have my cell on 902-8032.

NG - Update: 9:30am 'broke' into lab :D. Then found key...

11-2: VH, CZ
Nick ran the gel of I0500 restricited by Eco/Spe, but nothing showed up as expected (1.5kb).
Digested I0500 and J61003 with Eco/Pst as instructed.

2-5: MC, JG
ran gel of restricted I0500 & J61003; unfortunately there is no DNA in the I0500 sample either
gel extracted J61003
Made LB - to be divided into bottles & autoclaved in G308
O/N of DB in BB & DB in BE samples left in 37degree room

5-8: ML, ED autoclaved little bottles of LB. NOTE: does not need to be autoclaved before you pour it into little bottles and then re-autoclave. Just pour into bottles and autoclave once digested all in Boos except for DB with Pst/xba

to the top

September 10

Schedule: And Scheduling just got hard core

I have sheduled what we have taken down in the meeting some people appear many times on this schedule working alone just because of the time they were availible. If you are comfortable working alone and taking on lots of shifts im sure the team would be greatful but I'm also sure we would also understand if you didn't want to work alone in the lab. I have only scheduled this way because we need man hours so i tried to fill up all the spots as best i could. -JG

MC,JG for the AM
put away bottled LB
Ran gel of last night's restricted samples "in boo"; did not really turn out
ran gel of uncut & single restricted I0500; uncut I0500 showed 2 bands and restricted showed nothing
Mini prepped Ligations of DB in BB and DB in BE

to do: - run gel of double digested I0500
- do single & double restriction on Mini prep of Ligations
- run gel of uncut, single & double restrictions of mini prepped ligations

ML,AL for the MID (AM crew: What time will you guys be at the lab till? I won't be there until 12:30. Might need to arrange something in order to pick up keys. - AL)
Double digest of BB-DB/BE-DB XBA/PST

NG (2:30-4) for the PM (celine is in calgary if you dont feel comfortable in the lab by yourself I can show up - JG) Jason if you have time to show up that would be sweet, but if you don't I understand. -NG
Ran gel of the restrictions made in MID, Sept 10 by ML and AL.

JP,ED for the Night

to the top

September 11

Schedule: No one else really signed up for tuesdays so anyone who has time to drop in should

AM- NK Restrictions of BB+DB with XBA/PST and BE+DB with PST, SPE

Mid - AL (12:30 - 2 ish), NG (random)
Ran gel on restrictions made by NK in AM of Sept 11

Night- ED JP
Gel extractions
Ligations of DB+BB + BE and DB+BE + BB
mini relocation 1 bench back, do not use the first 3 benches of the lab or the blackboard from now on!

to the top

September 12


NG Hi guys I have a meeting 3:30 that cannot be reseached I moved myself to come in Tuesday instead (when nobody was in).

AM -

Mid- ML, AL

Important: Please evacuate lab before 2:00pm as there is a lab class starting at this time till 5pm! -AL

lab class 2-5 VH: please make note of the above announcement regarding lab class at 2:00pm

Night - CZ, ED
Transformed BEDBB and BBDB into XL 10 gold

to the top

September 13



Digested BEBOO and BBBOO with PST and XBA

Ran gel of restrictions made in the previous shift.

Meeting 7:00

After lab shift, JG, JP
Overnights with AMP of BBDB + BEDB
Gel extractions from gel made in the prevous shift
bands 1,2 (BE) correct while band 3 is too large to be BB
Sequence reactions can now be started using primers 1-5 that have been diluted to 5pmol/microlitre in H20 not TE

to the top

September 14


AM - MC (around 900/930hrs)
- Mini prep from O/N of BBDBE & BEDBB
- Glycerol stocks of BBDBE & BEDBB, and restreaked quartered plates
- Gel extraction of BE & BB Xba/Pst
- made gel

Mid - AL, JG
Restrictions on BBDBE and BEDBB with xba/pst and spe/pst
Ran gel on restrictions on BBDBE and BEDBB with xba/pst and spe/pst
Gel extracions

night - JP
Sequencing reactions
BBDBBE 6 reactions
BEDBBB 6 reactions
BB 3 reactions
BE 3 reactions

General note: Each primer sequence will be the end of the indicated gene to show us wether the joining/ligations worked correctly (VF is the promoter/gene/juction)
To do: Submit to MBSU

to the top

September 15


And yet another crazy weekend...

8-11 - ED,MC
General Note:We have cloned all 5 genes in series, in to differnt orders. Waiting for I0500 which couls be completed on monday. To make sure we have cloned correctly will also need to wait for the squencing results. This weekdn we will clone the 5 gense in 2 differnt orders. The digest have already been done
Gel extraction of gel bands of XBA,PST of BBDBBE and BEDBBB. THese can be used to cone into I0500 J61003
Ligations of DBBB into BE and DBBE into DBBEBB
Leave at 20 degrees celsius for 10 hours
5-8 - NK Cant find the competent cells.
Competent cells are in -80 freezer 2nd shelf from the bottom-ML, NG

to the top

September 16

Schedule: Continued...

8-11 ED, JG
Transform ligations of DBBEBB and DBBBBE into competent cells.
Plated on AMP with whole ligations
General Info: When DBBEBB and DBBBBE are complete run sequence reaction
Follow "flouresence sequence reaction" protocol

11-2 VH, CZ

2-5 MC, JP

5-8 - ML, NG
No growth on DBBBBE or DBBBBE at 1700
Retransformed into competent cells DBBEBB and DBBBBE
Plated 2 of each and lover overnight at 37 degrees

to the top

September 17

AM - JG (800 hrs - 930hrs),MC (@ 800hrs)
- delivered MSBU sequencing samples
- O/days of Ligations from yesterday

To do: miniprep, of DBBEBB and DBBBBE>
restrict with XBA, PST>
Seperate restriction SPE PST>
Ruun gel of UNCut ligations>
Cut with XBA PST and CUT with SPE PST>
GEl extract XBA, PST>

NB: AMP plates are marked with a red streak down the SIDE of the plate. if it does not have this - then it is NOT AMP

ML (~10:30-1:00)>

to the top

September 18

NK - 8 AM

VH - (@1pm)

AL - 12-2pm

to the top

September 19

ML - ~10:30 - 1:00

AL - ~1:00 - 2:00

Last Wednesday there was a lab class in our lab from 2-5, is this going to be a regular occurence? -VH


VH, see notes on Sept 12. - AL


to the top

September 20

NK - 8 AM

VH - (@1PM)

JG, MC- (@5PM)

JP- late

to the top

September 21

AM - MC, JG (@ 800hrs),

ML (~10:30 - 1:00)

AL - ~12:30 - 2:00

ED - 4 PM

to the top

September 22

ED 9-12

NG 12-3

NK 230-530

JP evening

to the top

September 23

ED 9-12

NG 12-3

NK 230-530

to the top

September 24

AM - MC @ 8000hrs

to the top

September 25

to the top

September 26

to the top

September 27

to the top

September 28

to the top

September 29

to the top

September 30

to the top

UofA iGEM Home
To August 2007