Schedule – week of Nov 3

Jack
1.  Vtg 1 promoter region
2.  Vtg Type 1 Std
3.  GST-pi Std
4.  Real Time
5.  Reverse transcription
6.  DNA stds for sybr green assay
June
Manuscript works and beta naphtaflovone exposure (if possible)

Emily
Study for prelims
Oversee processing of samples for RNA, histology and MC-LR analyses

Ping
Mon – setup spawning
Tues – collect embryos
Wed – setup spawning; WQ test
Thurs – collect embryos
Fri – water [...]

Lab outcomes – Oct 27-31

Mon

Setup spawning tanks
Paramecia checked
Larvae 102108 Day 6 – 100/beaker; the rest put in larvae box in big tank
Fed BS; no feeding observed
Larvae 102308 Day 4 – from petri dish to 1 beaker
Received permount

Tues

Spawning: ~600-800 embryos
All kept and labeled as 102808
No microinjection
Sonicated 100-200 micron particle larvae food
Fed to 102108 and 102308
Observed under microscope: seemed to be [...]

Schedule – week of Oct 27

Jack
1.  Vtg 1 promoter region
2.  Vtg Type 1 Std
3.  GST-pi Std
4.  Real Time
5.  Reverse transcription
6.  DNA stds for sybr green assay
June
Emily
Study for prelims
Oversee processing of samples for RNA, histology and MC-LR analyses

Ping
Mon – setup spawning
Tues – collect embryos
Wed – setup spawning; WQ test
Thurs – collect embryos
Fri – water change
*Monitor larvae growth
Ann
Candace
Jessica
Helping Jack
Rachel
Friday – Histology: Sectioning [...]

Lab outcomes Oct 20-24

Mon

Setup spawning tanks
Cleaned up lab
Paramecia checked

Tues

Spawning: ~500 embryos -labeled as 102108
Batch 6 all taken to be processed
Cleaned up lab

Wed

Setup spawning tanks
Cleaned up lab

Thurs

Spawning: ~380 embryos – labeled as 102308
~180 for microinjection
Water change in all tanks
Made sure everything is ready for inspection

Fri

Inspection
No histology – no permount

Microinjection Results

Candace Schaefer
Microinjection Report: 10/16/08
Submitted 10/20/08

1. Set-up

1.5% agar slants
Vortexed C60 soln 30 sec

C60 soln = 09/09/08 supernatant from 10 mg/ml C60 in DMSO

Centrifuged soln 15 sec
5% green C60

95 ul C60-DMSO
5 ul green food dye

Centrifuged 5% green C60 and DMSO (control) 10 min @ 10 rpm
Loaded needles
Set psi = 30 and broke needles

[...]

Schedule – week of Oct 20

Jack
Vtg 1 Promoter
GST-pi amplicon
B-actin std
Type 1 std
Real Time pcr

June
Manuscript, CYP1a1 preparation
Emily
Study for prelims
Oversee processing of samples for RNA, histology and MC-LR analyses

Ping
Mon – setup spawning
Tues – collect embryos
Wed – setup spawning; WQ test
Thurs – collect embryos
Fri – water change
*Monitor larvae growth
Ann
Experimental design write up ~2hrs
Microcystis
Weighing vials ~6hrs
Paramecia~3hrs
TOTAL ~11hrs
Candace
Jessica
RNA extraction – Emily’s samples
Rachel
Friday – Histology: Sectioning [...]

Lab outcomes – Oct 13-17

Mon

Setup spawning tanks
Batch 5 ready to go
Labelled cassettes
Paramecia checked

Tues

Spawning: ~500 embryos -labeled as 101408
No microinjection
Water change – big tank Nov/Dec Males & Females
Labelled cassettes

Wed

Setup spawning tanks
Prepared the remaining samples – Batch 6 723-860
Water change – medium tanks

Thurs

Spawning: ~700 embryos -discarded
~200 for microinjection
70% EtOH change in Batch 6
A few Oct males died

Fri

Histology

Lab outcomes – Oct 6-10

Mon

Setup spawning tanks
Changed 70% EtOH (Batch 4 #623-672)

Tues

Spawning: ~1000 embryos – discarded
No microinjection
Completed larvae growing box
Setup new dividers in big spawning tanks
Batch 4 taken to processor
Water change – 2 big and all medium tanks

Wed

Setup spawning tanks
Batch 3 received from processing
Prepared Batch 5 (#673-722)
Prepared cassettes

Thurs

Spawning: ~600 embryos – labeled as 100208
No microinjection

Fri

Spring break

Schedule – week of Oct 13

Jack
June
Emily
Study for prelims
Oversee processing of samples for RNA, histology and MC-LR analyses

Ping
Mon – setup spawning
Tues – collect embryos
Wed – setup spawning; WQ test
Thurs – collect embryos
Fri – water change
*Labelling cassettes; change out Bouin’s from vials; prepare samples to be sent for processing; monitor larvae growth
Ann
Candace
Wed = 2
+ set-up for injections
Thurs = 5
+ injections
Jessica
RNA extraction – [...]

Schedule – week of Oct 6

Jack
Mon – Pink Jeep Tour
Tues – hiking
Wed – grand canyon
Thurs – Antelope canyon
Fri – hiking
June
Away @ Chicago for conference
Emily
Study for prelims
Oversee processing of samples for RNA, histology and MC-LR analyses
Ping
Mon – setup spawning
Tues – collect embryos
Wed – setup spawning; WQ test
Thurs – collect embryos; water change
Fri -
*Labelling cassettes; change out Bouin’s from vials; prepare samples [...]