Lab outcomes – Aug 25-29

Mon

Ongoing experiment: PH#14
JP#3 started
Prepared for dissection on Thurs
Logged vials onto histology sheet

Tues

Spawning: ~450 embryos
Kept and labeled as 082608; not used for microinjection
Ended PH#14
Ongoing experiment: JP#3
Prepared for dissection on Thurs
Gave vials to John to take to ORNL

Wed

Ongoing experiment: JP#13
Setup spawning tanks
WQ test: pH 4.5 (Jan/Feb tank) -7.6; amm. 0.1-2.0 (Jan/Feb tank)
Changed water in tanks with [...]

Schedule – week of Sept 1

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 data analysis of JP Exp#3
Manuscript preparation of JP Exp#3
Preparation [...]

Schedule for week of Aug 25

Jack

ERE work
Linearizing transgene
Finish what I did not get done last week

June
JP Exp#3 exposure and termination
Emily

Go to JARTU 2x/day to administer salt treatment and feed fish
Catfish dissections on Thursday @ 12:30 (need Ping and Ann to assist)
Create spreadsheets and enter data from previous catfish dissections

Ping
Mon – setup spawning tanks; help June
Tues – collect embryos (saving [...]

Lab outcomes – Aug 18

Mon

Setup PH#13
Setup spawning tanks
Changed EtOH in vials (catfish project)
Updated RNA extraction SOP

Tues

Spawning: ~500 embryos
Kept and labeled as 082008; not used for microinjection
Ongoing experiment: PH#13
Prepared things needed for dissection on Wed
Changed EtOH in vials (catfish project)
WQ test: pH 7.9-8.1; amm. 0.1-1.4 (changed water in tanks with higher levels)

Wed

Ongoing experiment: PH#13
Setup 26 spawning tanks
Changed EtOH in [...]

Schedule for week of Aug 18

Jack
1.  Go blind b/c Ted wants to find ERE’s.
2.  Screen GST-pi clones/sequence
3.  Fluorescent scope
4.  Annoy Ping, Candace, and Ann as much as possible.
June
CYP1a1/EF-1 cloning
Experiment design
Emily

Go to JARTU 2x/day to administer salt treatment and feed catfish
Collect fish tissue samples on Lake Erie (17-19 Aug, dates subject to change depending on weather)
Catfish dissection for experiment on Wed [...]

Lab outcomes – Aug 11

Mon

Setup PH#11
Setup spawning tanks

Tues

Spawning:  ~250 embryos
Kept and labeled as 0801208; not used for microinjection
Ongoing experiment: PH#11
Prepared things needed for dissection on Wed

Wed

Ongoing experiment: PH#11
Setup spawning
Catfish dissection @ JARTU

Thurs

Spawning: ~750 embryos; labeled as 081408
Discarded 081208
Ongoing experiment: PH#11
Discussion – RNA storage
Prepare vials for Lake Erie Trip
Water change on medium tanks

Fri

Ended PH#11
Changed from Bouin’s solution into 70% ethanol [...]

Schedule for week of Aug 11

Jack

Finish prepping CFP and YFP for Candace to inject.
Candace and I will get with Polina to use Biggerstaffs fluorescent scope.
Screen GST-pi clones, DNA prep, In-Vitro transcription, quantification, dilutions, Real Time Std Curve.
Try to generate primers for Vtg 1 promoter.
Troubleshoot hsp27 promoter amplification. May pull another sequence from literature for amplification. The sequence [...]

Lab outcomes – week Aug 4

Mon

Medium tanks water change – performed on Sunday
Ongoing experiment: PH#8
Setup PH#10 (thanks to Jessica for helping)
Reported results of PH#9 Part 2 and PH#8
Received paramecia counting wheel

Tues

Spawning: ~1700 embryos
Kept and labeled as 080508; not used for microinjection
Ended PH#8
Ongoing experiment: PH#10
RNA extraction on PH#9 Part 2 by Jessica
Ammonia test – all good; values between 0.1-0.4 (from amm. [...]

Schedule for week of Aug 4

Jack
June
CYP1a1/EF-1 cloning
Experiment design
Emily

Go to JARTU 2x/day to administer salt treatment and feed catfish.
Prepare food for catfish experiment.
Prepare sample vials and gather supplies for catfish experiment.
Begin preparing sample vials and gathering supplies for Lake Erie trip (late August, dates TBD).
Revise methods section of manuscript.

Important dates for catfish experiment
(On each of these dates I will need [...]

Lab Outcomes – week July 28

Mon

Water change on all medium tanks
Setup PH#9 Part 2 & JP#3
Larvae raising – current count ~100
50 Larvae 072408 for Jack to do DNA extraction

Tues

Spawning: ~2200 eggs; ~150 for microinjection, saving the rest
Labeled as 072908
Ended PH#9 Part 1
Larvae raising – current count ~70 (disappointing)
BS did not hatch well today

Wed

Water change – glass bowls
Larvae raising – pathetic [...]