Difference between revisions of "User:Tara K. Luckau/Notebook/Team ConGen/2013/04/17"

From OpenWetWare
Jump to: navigation, search
(Autocreate 2013/04/17 Entry for User:Tara_K._Luckau/Notebook/Team_ConGen)
 
Line 9: Line 9:
  
  
==CNTI Acos5==
+
==SCOC MP2 State of the Multiplex==
  
  
===PCR===
+
===Summary of Past Results===
  
  
* comments
+
* SW614- A4, A6, B1, B12
:: [[Image:20130110_PCRa.png|800 px]]
+
** gelled well
 +
** amplified well in uniplex
 +
** amplified well in multiplex with all 8 SW markers
  
  
===Gel===
+
* multiplex with A4 + A6 + B1 + B12
 +
** gel blank
 +
** frag fail  ?!
 +
** maybe due to Techne cycler?
  
{| {{table}} style="text-align: center"
 
|-
 
! Pour !! Load !! Run
 
|-
 
| 270 mL 1x TAE + 5.4 g agarose || 2 µL gel load dye + 4 µL PCR product || 160 V
 
|-
 
| 27 µL GelRed || 6 µL ladder || 45 minutes
 
|}
 
  
 +
* re-do of multiplex with A4 + A6 + B1 + B12
 +
** (didn't run gel)
 +
** frag failed again!
 +
** ran on Techne again - thermalcycler poopy?
  
:: [[Image:20130110_Gela.png|800 px]]
 
  
 +
* re-re-do of multiplex with A4 + A6 + B1 + B12
 +
** gel blank
 +
** frag failed AGAIN!
 +
** ran on Opticon  :(
  
* mispriming everywhere - primer pair not usable for CNHY
 
* lots of mispriming, but may be able to use conditions indicated by orange face ([[Image:orangefrownyface.png|20 px]]), if needed, under stringent conditions
 
* favorable conditions indicated by [[Image:greenhappyface.png|25 px]]
 
  
 +
===Next Steps===
  
===Fragment Analysis Submission===
 
 
 
* UAGC Submission# ????
 
:: [[Image:20130227_Frag.png|900 px]]
 
 
 
* FedEx Tracking# [http://www.fedex.com/Tracking?language=english&cntry_code=us&tracknumbers=801124151779 801124151779]
 
:: [[Image:20130227_FedEx.png|700 px]]
 
 
 
* FedEx picked up ____pm, 26 February
 
* FedEx delivered ____am, 27 February
 
* UAGC received ____am, 27 February
 
* UAGC completed ____am, 28 February
 
  
 +
* redo it ALL to figure out what the problem is
 +
* potential problems:
 +
'''** DNA - run 4 samples, not just the 2 controls'''
 +
** fluorophore - can re-order labelled primers, but only after testing other options because of price
 +
** primer interactions - none to be concerned about
 +
** cycler - avoid using Techne until properly tested
 +
** 3730 - Crystal reports no problems with other clients' data
 +
'''** uniplex/4-plex/8-plex - just re-do these reactions'''
 +
** Taq - can run 2 replicates using 2 different lots of Taq, but don't know what lot I used for the above failures, so not a good test
 +
** Buffer - don't know how to test
  
  

Revision as of 14:41, 9 June 2013

Owwnotebook icon.png Tara's Lab Notebook <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>


SCOC MP2 State of the Multiplex

Summary of Past Results

  • SW614- A4, A6, B1, B12
    • gelled well
    • amplified well in uniplex
    • amplified well in multiplex with all 8 SW markers


  • multiplex with A4 + A6 + B1 + B12
    • gel blank
    • frag fail  ?!
    • maybe due to Techne cycler?


  • re-do of multiplex with A4 + A6 + B1 + B12
    • (didn't run gel)
    • frag failed again!
    • ran on Techne again - thermalcycler poopy?


  • re-re-do of multiplex with A4 + A6 + B1 + B12
    • gel blank
    • frag failed AGAIN!
    • ran on Opticon  :(


Next Steps

  • redo it ALL to figure out what the problem is
  • potential problems:

** DNA - run 4 samples, not just the 2 controls

    • fluorophore - can re-order labelled primers, but only after testing other options because of price
    • primer interactions - none to be concerned about
    • cycler - avoid using Techne until properly tested
    • 3730 - Crystal reports no problems with other clients' data

** uniplex/4-plex/8-plex - just re-do these reactions

    • Taq - can run 2 replicates using 2 different lots of Taq, but don't know what lot I used for the above failures, so not a good test
    • Buffer - don't know how to test