Klapperich Lab:Notebook/Lab Meeting Notes/2009/11/10

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10 November 2009 Lab Meeting

  • Attending:
  • Missing:
  • Presentation: Dr Shichu Huang from Auburn. 2pm, PLEASE BE ON TIME.
  • LUNCH at 12pm with Dr. Huang. Cathie's Treat. Please meet in at the 7th floor elevators. We will go to Bertucci's.

‡ Announcements

  • Oakridge deadline is 1 Feb 2010.
  • PAPERS Drafted before MicroTAS Are we done with this? 1st draft by Thur

‡ Flu R01:Integration

  • New design - address evaporation loss, Reagent delivery and storage solution. Consider marginating flows?
  • New design sent to make Rubber mold

† Sample Concentration (Lead: Jane, Team: Jaephil)

  • MDCK Cells are not propagating. Try troubleshooting efforts. Sonali will do on Monday.
  • Cassidy needs to see plaque assay again - so when cells are up, she needs training.
  • Up to high 10^7 for positive control. Silver substrate no signal.
  • Jane working on the cell lysate control.
  • Main loss is at the outlet/sample collection: tangential filtration design in drawing stage.
  • Contacted EC Shaw about rubber stamp mold for new design for evaporation.
  • Set up COMSOL and StarCD, look for governing equations for simulation of evaporation.
  • Committee meeting setup

† SPE Column Optimization for RNA. (Lead: Sonali, Team: Hussam, Jessie, Brendan)

  • Video with two color dyes and [red(primer) then blue, green, water+tween]
  • repeat experiment reported on 10/27 with 700nm Silica 3X.
  • controls for these: empties, non-silica monolith and full silica monolith, std. recipe.
  • Load DNA in a straw of 5,50,100ul SPE, quick test of hypothesis: more SPE = more DNA, To be repeated with Alex and Mark
  • Look into modifying Alex's pneumatic setup for the PATH guys. Instead of building the straw array over here NOTE: Sean
  • PLanning net silica per channel experiments. with 700nm, 3x possible. Deliver around 11/5/09.

  • New virus prep (JC, 6/11/09), getting Ct's of 27 for 1:16 dilution and 28 for 1:64 dilution. Use more diluted sample for future optimization experiments? Channels were 700nm silica/1X. pfu/ml is 10^6.
  • Tried 2 more chips with 3X 0.15um Silica. Chips are breaking b/c of no crosslinking. SPE was coming out even at 2ml/hr flow rate.
  • Running no silica and no SPE chips with virus. (JC/RNA) - No Silica channels grab comparable amounts of RNA with the channels with silica, lower with no SPE.
  • PCR is running on the samples from the new channel design. Will compare results with Hussam after the meeting and report back if significantly different.

† PCR - CMI (Lead: Qingqing)

  • QQ will work on the initial integration steps of SPE + RT (reservoir)+ PCR.
 - third design was tested with water,big bubbles observed, but the water still could be collected at the outlet.
 - third design was tested with PCR reagent(Flu assay). On-chip PCR does not work.
 - the same experiment was repeated with the reduced hot-start time(five minutes). the on-chip PCR still does not work.
 * New channel has been designed.
   - molds(SU-8,PDMS,Epox)has been made.
   - chip has been made and tested with water. Both thermal and fluidic control are fine. it will be tested with PCR reagent this week.
  • PCR of C.Difficile DNA
 - primer for toxin A.
    - on chip PCR with the optimized primer and template concentration   does not work.
    - MgCl2 concentration was optimized to improve the PCR efficiency.
    - on chip PCR with the optimized MgCl2 concentration does not work.
 - Primer for toxin B.
   New primer sequence from Lisa has been ordered, will be tested when they come.
  • PCR2 Paper formatted for LOAC this week.
 need to get back from Cathie
  • CMK: PCR 1 draft. Analytical Chem. MCK is running more simulations.

† HDA (Lead: Jaephil, Team:Sonali)

  • Start planning R01 for Submission on 2/5/10. MM, JD, CMK.
  • HDA chip fabrication training will be after new process settled (comeback from mTAS).
  • Two methods, Hot embossing and cutter plotting will be combined to reduce whole process time. New mold design sent.
  • Paper submitted 10/6.

‡C. diff Project (Cathie, Sonali, Satish Singh, Lisa J., His post doc )

  • Sonali to train Lisa on SPE.
  • QQ to run test PCR on chip with genomic DNA and Toxin B primers.

‡ Coulter Flu Fraunhofer Project (Lead: Sonali, Team: Sonali, Jessie, Cathie, CMI Folks, Qingqing)

  • Meeting 9am Monday (every other week)

‡ Agilent Automated Sample Preparation (Lead: Alex)

  • Bacillus subtilis gDNA and cells were tested on SNAP2 machine. gDNA gave good results, pcr signals from cell-samples still low. More tests ongoing. Hot Dog samples will follow.
  • First Draft of Yeast Paper send to Cathie and Alexis.
  • Nano-C: which cells shall we use for lysing experiments?

‡ COBRA (Lead: Jaephil, Team: Cathie, Jane for virus only, PHO folks)

  • Metal piece modification for the integration - Fix did not turned out good. Make new or try fix again?
  • paper 1, 1st draft by Thu(10.29): Evap with Sol Gel substrate. Not integrated. MSSA, E coli.

‡ Biointerfaces group (Lead: MinCheol, Team: Cathie, MCK, Wong and Meller folks)

  • New experiments happening now.
  • Cathie will submit paper inquiry.

‡ CIMIT- Sepsis (Lead:Cathie, Team:TBA)

  • IRB approved.
  • Cathie submitted the companion BU IRB form for exemption. Revised, still still waiting.

‡ PATH Grant (Lead:Cathie, Team:Frank, Sean, Mark, Jake Trueb (ME), Suma)

  • Phone meeting with PATH this week. 10/13.
  • Senior Project Proposals due in less than two weeks! (23 Nov.)
  • Frank: 1-week unstabilized sample PCRed (increased degradation with higher storage temps as expected; eluting 2-week sample on Wed.
  • Frank: PVA to be ordered for home-made polymer storage solution; begin solubility tests upon arrival.
  • Sean: Drawings with measurements have been made. Currently working on learning CAD software and making a professional CAD drawing.
  • Sean met with Alex for an integration of straws on black machine. They sketched a design. Sean will make a professional drawing and create a required parts list. Alex will supervise the design. Before handing in the job to the shop floor, the design will be presented to Cathie.
  • Mark to begin initial DNA testing with shorter time periods (1,3 days, etc.) to be completed next week. PCR training to be completed with Hussam.