User:Michael F. Nagle/Notebook/Chem 571/2012/10/16

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
Line 8: Line 8:
==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
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

Revision as of 01:45, 7 December 2012

Project name Main project page
Previous entry      Next entry

Procedure

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

Image: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


Personal tools