Team:HokkaidoU Japan/Notebook/September8

From 2010.igem.org

(Difference between revisions)
 
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=Confirmation of pSB1C3=
=Confirmation of pSB1C3=
Because pSB1C3 PCRed from 1-3A(RFP reporter) was used there was posibility of contamination by template which would also produce red colonies.
Because pSB1C3 PCRed from 1-3A(RFP reporter) was used there was posibility of contamination by template which would also produce red colonies.
-
* H buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation
+
* H buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → [[Team:HokkaidoU_Japan/Protocols|Transformation]]
* M buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation
* M buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation
-
* H buffer treated 2 uL → [[Team:HokkaidoU_Japan/Protocols|Transformation]]
 
* H buffer treated 2 uL → Transformation
* H buffer treated 2 uL → Transformation
 +
* M buffer treated 2 uL → Transformation
=PCR of GFP=
=PCR of GFP=

Latest revision as of 08:14, 27 October 2010

Colony PCR

Checked if primers PS-R, EX-F were good to use for colony PCR and also checked if insert was realy in plasmids

  • September 6th transformation (digestion was done with H buffer×2 and M buffer×2, 2 version)
  • 1-3A
  • Retried September 6th transformation
  • pUC119

Confirmation of pSB1C3

Because pSB1C3 PCRed from 1-3A(RFP reporter) was used there was posibility of contamination by template which would also produce red colonies.

  • H buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation
  • M buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation
  • H buffer treated 2 uL → Transformation
  • M buffer treated 2 uL → Transformation

PCR of GFP

  • GFP reporter BBa_I13522 pSB1A2 2-8A 937 bp
  • GFP protein BBa_E0840 pSB1A2 1-12O 878 bp
PCR cocktail was same as September 1st's