IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/30: Difference between revisions

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==Entry title==
==Team Allergy==
* Insert your content here.
*Sequencing Results:
LTP ihpRNA parts that worked: 9c2, 11c1, 11c2


Bet 2 ihpRNA parts that worked: 25c1,c2, 28c1,(LTPS+PDK is actually Bet 2 hpRNA)
Ger ihpRNA parts that worked: 36c1,c2, 37c1,2, 38c1,c2
==Team Flavor==
* Ligated StrepII tagged proteins into V24 backbone
* Ligated cut J45004 into V0120 backbone
Ligations were plated and left @ 37°C O/N.


==Team Fence==
==Team Fence==


===PCR of VP16 and Gal4DBD===
===PCR of VP16 and Gal4DBD===
{| class="wikitable" border ="1px solid black"
|-
!1
!2
!3
!4
!5
!6
!7
!8
!9
!10
!11
!12
|-
|
|KB+ Ladder
|Gal4 DBD 1
|Gal4 DBD 2
|Gal4 DBD 3
|Gal4 DBD 4
|VP16 1
|VP16 2
|VP16 3
|VP16 4
|KB+ Ladder
|
|-
|}
[[Image:PCR_VP16_Gal4DBD_7-30.jpg]]
[[Image:PCR_VP16_Gal4DBD_7-30.jpg]]


===PCR again for VP16===
Using pfx polymerase
*1μL 1/10 dilution Hind.Fwd
*1μL 1/10 dilution VP16.Rev
*1μL DNTPs
*6μL enhancer buffer
*.5μL MgSO<sub>4</sub>
*.5μL PFX polymerase
*1μL 1/10 dilution VP16 #1
*5μL DH<sub>2</sub>O
*4μL amp(lification?) buffer
Running PCR on two reactions. There was not enough PFX for both to get .5μL, so only one of them is likely to actually amplify.
===Second ACC + RxrHm digest gel===
[[Image:sucesspcrkfkdigestacvtually.jpg]]
yes to ACC (left) no RxrHm
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Revision as of 15:29, 1 August 2010

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Team Allergy

  • Sequencing Results:

LTP ihpRNA parts that worked: 9c2, 11c1, 11c2

Bet 2 ihpRNA parts that worked: 25c1,c2, 28c1,(LTPS+PDK is actually Bet 2 hpRNA)

Ger ihpRNA parts that worked: 36c1,c2, 37c1,2, 38c1,c2

Team Flavor

  • Ligated StrepII tagged proteins into V24 backbone
  • Ligated cut J45004 into V0120 backbone

Ligations were plated and left @ 37°C O/N.

Team Fence

PCR of VP16 and Gal4DBD

1 2 3 4 5 6 7 8 9 10 11 12
KB+ Ladder Gal4 DBD 1 Gal4 DBD 2 Gal4 DBD 3 Gal4 DBD 4 VP16 1 VP16 2 VP16 3 VP16 4 KB+ Ladder

PCR again for VP16

Using pfx polymerase

  • 1μL 1/10 dilution Hind.Fwd
  • 1μL 1/10 dilution VP16.Rev
  • 1μL DNTPs
  • 6μL enhancer buffer
  • .5μL MgSO4
  • .5μL PFX polymerase
  • 1μL 1/10 dilution VP16 #1
  • 5μL DH2O
  • 4μL amp(lification?) buffer

Running PCR on two reactions. There was not enough PFX for both to get .5μL, so only one of them is likely to actually amplify.

Second ACC + RxrHm digest gel

yes to ACC (left) no RxrHm



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