User:Jose Fabricio Lopez/Notebook/Logbook/2011/08/18: Difference between revisions

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==PCR of pSB1T3==
==PCR of pSB1T3==
 
*PCR of pSB1T3/RFP, to amplify just backboney
 
{| border="1"
!Reactives
!μL
|--
|Buffer Taq Platinum 10X
|5
|--
|dNTPs 8mM
|--
|MgCl
|1.5
|--
|1.5
|--
|Oligo SuffixF/PrefixR
| 2/2
|--
| DNA [[Jose Fabricio Lopez/Notebook/Logbook/2011/06/19|pSB1T3]]
|1
|--
|H<sub>2</sub>O
|33
|--
| Taq Platinum DNA Pol
| .5
|--
|Total
| 50μL
|}




Revision as of 11:24, 19 August 2011

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Objectives

  • PCR of B0015.
  • PCR of pSB1T3.
  • See the in Gel.

Working

PCR of B0015

The point of this PCR is get double terminator and insert it into pSB1T3 backbone.

  • Amplified with preffix and suffix primers.


Reactives μL
Buffer Taq Standard 10X 5
dNTPs 8mM 1.25
Oligo F/R 2.5/2.5
DNA B0015 1
H2O 36.75
Taq DNA 1
Total 50μL

Gel B0015

  • Lane 1. Ladder
  • Lane 2 & 3. B0015

It's dirty but usefull

PCR of pSB1T3

  • PCR of pSB1T3/RFP, to amplify just backboney
Reactives μL
Buffer Taq Platinum 10X 5
dNTPs 8mM
MgCl 1.5
1.5
Oligo SuffixF/PrefixR 2/2
DNA pSB1T3 1
H2O 33
Taq Platinum DNA Pol .5
Total 50μL



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