Lab Notebook - Elicia

 Notebooks Home Page

6-22-09
Followed Protocol( Revisions were made for a triple cell content) Each well had 200 uL of the cells Check OD Measurements Strep Trial 1

Last 4 Wells No Wash- top



Last 4 Wells-bottom Flipped upside down no wash 5uL of strep/PBS (1 part to a part was added to the wells Incubation for 45 min Cells were spinned down (followed protocol)



(Last 4 Wells top Wells A 9-12 and B12 (washed only)) Cells Wells A 9-12 and B12 were washed with 100 uL PBS

(Last four Wells-top Wash) All Wells were washed with 100 uL PBS (Wells A 9-12 and B12 were not rewashed)

Analysis of Results Not what we wanted to see Well C-12 supernatant was still in the well before PBS was added?

What we will do…. Revise the strep protocol Use a new positive control pBca 9145- Bca 9494

6-23-09
diluted strep plasmids

6-24-09
Bio277

Analytical Gel

induced strep plasmids. Parts and vector(??) were digested

6-25-09
Did ligation(vector used ??), transformation(used TG1 cells- plated on amp)and strep

6-26-09
Redid ligation(used pBca 9523-1144 #5) and transformation (used TG1 cells plated on Spec and did heat shock) ....awaiting results (keeping fingers crossed) ....awaiting new streptavin to arrive on Monday June 29

6-28-09
Plates are beautiful- no red and big white cells are present. Annoculated (+) and (-) controls for Strep (oh no induced overnight and did a 2% concentration- that's not right always do a 1% and induce in the morning so it can be used in 3-6 hours- was redid by Tom)

6-29-09
Mini Prep (used zymo columns- must redo)

6-30-09
Redid Mini Prep

Analytical Gel Send parts for Sequencing yayyy...

7-3-09
Reaequenced Bioig 277 and 324



Sent for Sequencing Eig55 and 56

7-6-09
bowling

7-8-09
Poured spec and amp plates. Autoclaved glassware. Set up strepspread sheet.

7-24-09
K11212 that was sent off for sequencing

7-25-09
Mini prep 24- well plate samples. Only Bdr0ll-2(Aa),Bdr0ll-1 #2(Ba),MV45-2(Ca), Bdr0ll-2#2(Da), Bdr0ll-1#3(Ea),Bdr0ll-2 #3(Fa),Bdr0ll-1#1(Ga)

Order is:Bdr0ll-2(Aa),Bdr0ll-1 #2(Ba),MV45-2(Ca), Bdr0ll-2#2(Da), Bdr0ll-1#3(Ea),Bdr0ll-2 #3(Fa),Bdr0ll-1#1(Ga) Colony PCR from Plates MV45-1(Ab), 009-1(Bb),k11212-2(Cb), 008-1(Db),009-2(Eb), MV45-2(Fb), k112126-1(Gb), k112126-2(Hb), k11212-1(Ib),008-2(Jb) Order is:MV45-1(Ab), MV45-1(Ab),009-1(Bb),009-1(Bb),k11212-2(Cb),k11212-2(Cb), 008-1(Db),Nothing, Marker, 009-2(Eb), 009-2(Eb),MV45-2(Fb), MV45-2(Fb),k112126-1(Gb),k112126-1(Gb), k112126-2(Hb), k112126-2(Hb),k11212-1(Ib),k11212-1(Ib),008-2(Jb),008-2(Jb)

7-26-09
Redid PCR k112126-1(Ac),k112126-1(Bc),008-1(Cc),008-1(Dc),009-1(Ec),-009-2(Fc),MV45-1(Gc),MV45-2(Hc),k11212-1(Ic),k11212-2(Jc)

7-27-09
k112126-1(Ac),k112126-1(Ac),k112126-1(Bc),k112126-1(Bc),008-1(Dc),008-1(Dc),009-1(Ec),009-1(Ec),009-2(Fc),009-2(Fc),MV45-1(Gc),MV45-1(Gc),MV45-2(Hc),MV45-2(Hc),k11212-1(Ic),k11212-1(Ic),k11212-2(Jc)k11212-1(Ic),(008-1 Cc turned pink- so it was not included in this gel) Grew in LB AMp:k112126-1(Ac),k112126-1(Bc),008-1(Cc),008-1(Dc),009-1(Ec),-009-2(Fc),MV45-1(Gc),MV45-2(Hc),k11212-1(Ic),k11212-2(Jc)

7-28-09
Miniprep Fig3 parts only k112126-1(Ad),k112126-1(Bd),008-1(Dd),MV45-2(Hd),k11212-1(Id),k11212-2(Jd) Gc,Ec: did not grow Fc:turned pink

.