Friday, May 22, 2015

Direct PCR for the insert clones

5/6/2015
Clones were grown on LB+ Hygromycin 250 mg/ml, direct sample was taken from the colony.

  • Clones: ClpX1, X2, X3, X4, X5, X6, X7,X8 ,X9 ,X10 and X11  
  • ClpX primar with pJH1 clone as a control.
  • ClpX PCR as a positive control.

  • ClpB1, B2,B3, B4, B5, B6, B7and B8 .
  • ClpB primar with pJH1 clone as a control.
  • ClpB PCR as a positive control.

  • ChiT1,T2,T3 and T4 
  •  ChiT primar with pJH1 clone as a control,ChiT primar with B/r E. coli clone as a control. 
  • ChiT PCR as a positive control.



1  μL      3` R primares
1  μL      5` F primares
12.5 μL   supermix.
10 μL dd water.

Protocol:
 95 °C    3 minutes

95 °C   30 seconds
55 °C  2 minutes                    30 cycles
68 °C  1.5 minutes

Final elongation 68 °C  for 5 minutes.




ChiT B/r
ChiT,pJH1
ChiT PCR 
ChiT3

ChiT2
ChiT1



ClpB,pJH1
ClpB,PCR
ClpB3
ClpB2
ClpB1
ClpX,pJH1
ClpX,PCR
ClpX3
ClpX2
ClpX1

ladder

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