Lab outcomes – week Sept 22-26

Mon

Setup spawning tanks
Labelled cassettes
Prepared tissue for processing (#501-512, 530-565 –> Batch 2) – ready to go
Labelled vials/tubes for last catfish dissection

Tues

Spawning: ~700 embryos – all discarded
No microinjection
Discarded 091808
Weighed vials for catfish dissection
Liver,muscle and brain dissection @ JARTU
Labelled cassettes
Prepared next batch of samples (#573-622 –> Batch 3) for processing
Made dividers for big spawning tanks

Wed

Setup spawning tanks
Vials [...]

Schedule – week of Sept 29

Jack
1. continue with vtg 1 promoter
2. confirm GST-pi
3. in-vitro transcription of type 1 amplicon
June

Manuscript preparation
Preparation for presentation

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 – inspection/tour of facility
*Labelling cassettes; change out Bouin’s [...]

Candace’s microinjection results

DiI:DMSO (1:4)
 

well

20-50 ms

100 ms

200 ms

500 ms

F1

F2

 

 

 

 

F3

 

72 hpf
(same fish 3 images below)
 
 

F4

 

 

 

F5

 

 

72 hpf
(malformed)

F6

 

 

72 hpf
 

F8

 

 

 

72 hpf

500 ms
 
          

 
 

Candace’s microinjection results

Candace Schaefer
Solutions 09/12/08
DiI-EtOH in PBS (1:1)
50 ms

DiI-EtOH in PBS (1:1)

DiI-EtOH in DMSO
1:1?
5ms                     5 ms (bright)
1:2?
25 ms                    5 ms (bright)
1:4
25 ms
1:9
25 ms                25 ms (bright)            25 ms (bright)
DiI-EtOH in C60-DMSO
1:4
25 ms                25 ms                      25 ms        [...]

Lab outcomes – Sept 15-19

Mon

Setup spawning tanks
Dissection @ JARTU

Tues

Spawning: ~600 embryos; labeled as 091608
~150 for microinjection
Labeled tubes

Wed

Setup spawning tanks
Vials labelling
Received processed liver tissue samples
Record vials into histology log sheet
June prepared C60 solution for Shaun

Thurs

Spawning: ~600 embryos; labeled as 091808
~150 used for microinjection
Discarded 091608
Water change – medium tanks
DIssection @ JARTU

Fri

WQ test
Changed Bouin’s solution to 70% EtOH from Thurs’ dissection
Histology
Vials/tube [...]

Schedule – week of Sept 22

Jack
1. vtg 1 Promoter – experimental design and hopefully PCR
2. RTPCR of PH11,12,13,confirm some of PH1 results, and Candaces samples.
3. GST-pi – sequence, in-vitro transcription, make std curve, run RTPCR.
4. Zfin clone came, amplify vtg 1 promoter from it.
June

Manuscript preparation
Preparation for presentation

Emily
Ping
Mon – setup spawning
Tues – collect embryos
Wed – setup spawning
Thurs – [...]

Lab outcomes – Sept 8-14

Mon

Setup spawning tanks
RNA extraction
Prepare cassettes
Paramecia flasks split

Tues

Spawning: ~500 embryos; labeled as 090908
~150 for microinjection
Labeled tubes
WQ test: pH 7.5-7.9; amm. level 0.1-0.3
Received 12 new 400mL beakers

Wed

Setup spawning tanks
Looked up powdered larvae food
Vials labelling

Thurs

No embryos – light was on the whole night
Obtained liquid nitrogen
Setup spawning tanks with Oct fish

Fri

Spawning:
~150 used for microinjection
Water change – medium tanks

Sat [...]

Schedule – week of Sept 15

Jack
June

Vit gene expression data analaysis
Manuscript preparation
Preparation for presentation

Emily
Go to JARTU 2x/day to salt and feed catfish
Catfish dissections on Monday and Thursday
Study for prelims
Ping
Mon – setup spawning; dissection @ JARTU
Tues – collect embryos
Wed – setup spawning
Thurs – collect embryos; dissection @ JARTU
Fri – WQ test; water change
*Prepare for dissection; change out Bouin’s from vials; record vials [...]

Lab outcomes – Sept 1-5

Mon

Labor Day Holiday

Tues

Setup spawning tanks
Prepared for next week’s dissection

Wed

Spawning: ~1000 embryos; labeled as 090308
~150 for microinjection
Setup spawning tanks
Labelling vials
Prepared cassettes

Thurs

Spawning: ~1000 embryos; labeled as 090408
Kept and labeled as 0828008; ~150 used for microinjection
Prepared vials
Prepared cassettes

Fri

Water change – medium tanks
RNA extraction – JP#3
LMB RNA extraction

Schedule – week of Sept 8

Jack
1. vtg 1 Promoter – experimental design and hopefully PCR
2. RTPCR of PH11,12,13,confirm some of PH1 results, and Candaces samples. Do all these need all 3 types of vtg and all controls?
3. GST-pi – sequence, in-vitro transcription, make std curve, run RTPCR.
June

Vit gene expression analaysis
Manuscript preparation
Preparation for presentation

Emily
Training for the catfish [...]