Alberta/Calender/August
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August 1
Schedule: CZ,NG,VH
General Notes:
1. Run gel of B0034 SpeI/PstI double digest; extract gel with QIAgen kit; stored in freezer.
2. PCR for
a. Enny + B0034 miniprep 1 to 8
b. Betty colonies (10)
c. Buddy colonies (10)
Follow James' protocol. Also see blackboard for James' comment about PCR.
For tomorrow
1. run gel of PCR prodcut which are in the thermocycler �
2. check restreaked single colonies of the Betty and Buddy colony PCR
3. grow up Betty and Buddy overnight and for miniprep and ligation later
4. Make Amp plates (we only have 4 more)
5. Autoclave test tubes
Quotes of the day: We are tired.
August 2
Modeling Meeting @ 1830hrs in CAB373
General Meeting @ 1900hrs in CAB373
Party afterward's @ Garneau pub
Schedule:
MC, JP & AF are meeting at 1700hrs to get some stuff done before the meeting happens; come join in the fun if you'd like.
Lab Notes:
Made AMP plates
Autoclaved test tubes
Ran gel of ?
Started O/N of Buddy & Betty
Checked restreaked colonies of Buddy & Betty --> positive growth except for 9&10 but this is expected because they are controls
James did a loving PCR of Benny in B0034
-MC, ED, JP, JB
August 3
Schedule:
Nik cant make it he just isn't crazy enough looks like justin is currently the only one crazy enough
Michelle & Justin are now the long weekend crew - be prepared for ILLUSIONS!!!! or perhaps just some awesome pipetting techniques while they're prepping to champ out at the ULTIMATE pipetting Olympiads.
Meeting @ 1700hrs
Captain's Log, Stardate: 080307-1153
Minipreps of lifeforms Buddy and Betty
Not enough room in box with blue tape, designed a new holding vessel (i.e. box with white tape below box with blue tape in the freezer) for minipreps of aforementioned lifeforms.
-JB
Made gel
Digested (double) Buddy & Betty minipreps
Digest (single) I0500 - from tube we believe is a miniprep in the -20 freezer
weekend to-do list
-JP, ML, MC
August 4
Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030
Lab Notes:
Made a project timeline - check out the cabinets on the left side of the 4C-degree fridge
Ran gel - no bands for I0500 (we were sad) but there are awesome bands of Betty & Buddy
Attempting to figure out the I0500 problem; transformed into XL10 Gold & HB101
-MC, JP, ML
August 5
Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030
Lab Notes:
1-transform I0500 into HB101 and XL10 gold
2-restricted mini's of Buddy and Betty with Xba/Pst (confirmed with gel their presence yesterday)
3-ran I0500 samples in gel and no DNA present, chucked out samples - re transform...
-MC, JP & ML (in spirit)
For Tomorrow
1-gel purify Buddy and Betty
2-ligate Buddy and Betty into Boo34 (overnite)
3-start ONs of I0500 for mini on tuesday
August 6
Meeting at 1000hrs - Michelle will definately be there at 1000 Justin is expected at 1030
Lab Notes:
- JP, ML, AF, MC
Notes:
1- re-plated I0500 tranformations on Kan and AMP plates - hope for growth
2- ran buddy and betty XbaI/PstI restriction products on gel and gel extracted
3- began ligation reaction with B0034 (in back shaker at 13 degrees)
4- started making butanol plates...but it might not work so well with higher percentages because when you plate the agar its warm/hot and when you add the butanol it bubbles like mad (evaporates). We tried pouring at lower temp, but it prooved disasterous
Fact of the Day
A good ligation is like a hoola-hoop, they're both circular and only one is methylated
August 7
Schedule:
MC,VH,AF
Protocol for Today
1: If there is growth on I0500 plates in 37degrees, make some overnites of 5 colonies (4ml LB + 2.4microL KAN [in buffer box]in shaker tubes and innoculate with bacterial colony and place in shaker at 37degrees overnite -circle each colony you pick and match with respectively labeled shaker tube)
2: Take the two ligation mixtures from the back room shaker- transform into XL10Gold bacteria- for each of the sample make two dilutions.
dilute (1)2microL into 8microL water and
(2)1microL into 9microL water.
Take the four samples and transform them (dilute in 600microL LB before puting into shaker.
Plate each transformant onto AMP plates in two dilutions:
(1)200microL on a plate
(2)50microL + 50microL LB.
Lab Notes:
No growth on Kan plates of I0500 in HB101 - therefore no O/N
Transformations of Buddy+B0034 Ligations & Betty+B0034 Ligations into XL10GOLD competent cells
- Betty+BOO34 10x = 1ul Lig + 9ul H20 (the number on the side of the eppendorf denotes the ratio of vector to insert in the lab book) 20x = 2ul Lig + 8ul H20
- labelling same for Buddy+BOO34
- MC, VH, AF
August 8
Today is sweatpants day. Wear your favorite pair.
The last of the synthetic genes has arrived (butyraldehyde dehydrogenase = BuAld_DH). I resuspended the plasmid in 40uL TE and put the red capped tube (with a pink tape flag) in the freezer in G308 - MKD
Schedule:
ED,NK,JP
August 9
Schedule:
ED,AF,JG
August 10
Schedule:
CZ, MC, JG
would anyone be able to take my spot??? thanks - MC
I'll let ya know tomorrow evening if i can
Modeling meeting cancelled as Nick G. and Doug are both away - WM
August 11
Schedule:
JP, AL, NK
August 12
Schedule:
CZ, AL, NK
August 13
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