User:Tara K. Luckau/Notebook/Team ConGen/2011/06/23: Difference between revisions

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==MP1==
==SCOC Multiplex 1==




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* similar to 13 June 2011 PCR - uniplexes of each primer pair, then multiplex, using labeled primers
* similar to 13 June 2011 PCR - uniplexes of each primer pair, then multiplex, using labeled primers
* [[Image:20110623 PCR.jpg|800 px]]
* [[Image:20110623 PCR.jpg|800 px]]
===Gel===
{| {{table}} style="text-align: center"
|-
! Pour !! Load !! Run
|-
| 270 mL 1x TAE + 5.4 g agarose || 1 µL gel load dye + 5 µL PCR product || 160 V
|-
| 27 µL GelRed || 6 µL ladder || 50 minutes
|}
* see 13 June 2011 for banding pattern expectations
* [[Image:20110623 Gel.jpg|800 px]]
* consistent with 13 June 2011 results (missing Scun22 band, but all others seem to be there)
* notice bright primer band on multiplex only - probably could do some optimization to make reaction more efficient
==SCOC Multiplex 1==
* SCOC Multiplex 1 diversity panel with labelled primers to send to frag, part deux!
===PCR===
* [[Image:20110623 PCR2.jpg|800 px]]
* some of the samples used for the diversity panel are getting low ... should switch out soon
* SCOC TP3 X01 was low, like the others, but I already had a SCOC TP3 extracted last year in the Mass Extract ... so used it, SCOC TP3 X06




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* see 13 June 2011 for banding pattern expectations
* see 13 June 2011 for banding pattern expectations
* [[Image:20110623 Gel.jpg|800 px]]
* [[Image:20110623 Gel2.jpg|600 px]]
* a whole lotta nada  :(
* still a whole lotta nada  :(
* strong primer band, again
* a bit of amplification of Scun9/10/11, but it's crap
 


===Next Steps===
* try full gradient on multiplex, ugh




Revision as of 09:28, 24 June 2011

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SCOC Multiplex 1

PCR

  • similar to 13 June 2011 PCR - uniplexes of each primer pair, then multiplex, using labeled primers


Gel

Pour Load Run
270 mL 1x TAE + 5.4 g agarose 1 µL gel load dye + 5 µL PCR product 160 V
27 µL GelRed 6 µL ladder 50 minutes


  • see 13 June 2011 for banding pattern expectations
  • consistent with 13 June 2011 results (missing Scun22 band, but all others seem to be there)
  • notice bright primer band on multiplex only - probably could do some optimization to make reaction more efficient


SCOC Multiplex 1

  • SCOC Multiplex 1 diversity panel with labelled primers to send to frag, part deux!


PCR

  • some of the samples used for the diversity panel are getting low ... should switch out soon
  • SCOC TP3 X01 was low, like the others, but I already had a SCOC TP3 extracted last year in the Mass Extract ... so used it, SCOC TP3 X06


Gel

  • used left-over gel from 13 June 2011
Load Run
2 µL gel load dye + 4 µL PCR product 160 V
6 µL ladder 1 hour


  • see 13 June 2011 for banding pattern expectations
  • still a whole lotta nada  :(
  • strong primer band, again
  • a bit of amplification of Scun9/10/11, but it's crap


Next Steps

  • try full gradient on multiplex, ugh



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