Difference between revisions of "User:Michael F. Nagle/Notebook/Chem 571/2012/10/16"

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==Procedure==
 
==Procedure==
 
* The procedure for [[AU Biomaterials Design Lab:Protocols/PCR Mutation Protocol|PCR Mutation]] was followed  
 
* The procedure for [[AU Biomaterials Design Lab:Protocols/PCR Mutation Protocol|PCR Mutation]] was followed  
** The specifications of the primer used are as follows:
+
** The specifications of the forward primer used are as follows:
 
[[Image:Primer212.png]]
 
[[Image:Primer212.png]]
 
**1mL molecular biology grade water was added to the primer.
 
**1mL molecular biology grade water was added to the primer.
 
**The volume of primer solution needed to prepare 1mL of 100ng/uL solution was calculated
 
**The volume of primer solution needed to prepare 1mL of 100ng/uL solution was calculated
Target concentration: 100ng/uL
 
 
***0.49mg/1000μL = 490ng/μL
 
***0.49mg/1000μL = 490ng/μL
 
***100ng/uL*1000uL / 490ng/uL = 204.08μL
 
***100ng/uL*1000uL / 490ng/uL = 204.08μL
 +
**[[User:Keyun_Wang/Notebook/Experimental_Biological_Chemistry_I/2012/10/16|Keyun Wang]] prepared 100ng/uL solution of the matching forward primer.
 +
**Solution was prepared with the following:
 +
***1uL forward primer ***1uL reverse primer ***1uL wild strand ADA ***5uL 10x Pfu buffer ***40.6uL molecular biology grade water ***1uL Turbo DNA polymerase solution ***0.4Turbo DNA polymerase 25mM dNTPs
  
 
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Revision as of 23:45, 6 December 2012

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Procedure

  • The procedure for PCR Mutation was followed
    • The specifications of the forward primer used are as follows:

Primer212.png

    • 1mL molecular biology grade water was added to the primer.
    • The volume of primer solution needed to prepare 1mL of 100ng/uL solution was calculated
      • 0.49mg/1000μL = 490ng/μL
      • 100ng/uL*1000uL / 490ng/uL = 204.08μL
    • Keyun Wang prepared 100ng/uL solution of the matching forward primer.
    • Solution was prepared with the following:
      • 1uL forward primer ***1uL reverse primer ***1uL wild strand ADA ***5uL 10x Pfu buffer ***40.6uL molecular biology grade water ***1uL Turbo DNA polymerase solution ***0.4Turbo DNA polymerase 25mM dNTPs