User:Kathryn Muratore/Notebook/AU CHEM-570 lab prep/2011/06/16: Difference between revisions

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|34.5 μL ||5 μL ||0.5 μL ||5 μL of 82 μg/mL ||5μL
|34.5 μL ||5 μL ||0.5 μL ||5 μL of 82 μg/mL ||5μL
|-
|-
![[User:Kathryn_Muratore/Notebook/AU_CHEM-570_lab_prep/2011/06/13| BSA PCR]]
![[User:Kathryn_Muratore/Notebook/AU_CHEM-570_lab_prep/2011/05/26| BSA PCR]]
|24.5 μL ||5 μL ||0.5 μL ||15 μL directly from PCR reaction ||5μL
|24.5 μL ||5 μL ||0.5 μL ||15 μL directly from PCR reaction ||5μL
|}
|}
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*→ 20' @ 65°C (it took awhile to get the block to the right temp, so they were on ice for a bit, then at ~70°C before the block reached 65°C)
*→ 20' @ 65°C (it took awhile to get the block to the right temp, so they were on ice for a bit, then at ~70°C before the block reached 65°C)
* Note: I should have done a PCR cleanup on the BSA PCR sample! I will repeat this. *'''[[User:Kathryn Muratore|Kathryn Muratore]] 10:20, 20 June 2011 (EDT)''':
* Note: I should have done a PCR cleanup on the BSA PCR sample! I will repeat this. *'''[[User:Kathryn Muratore|Kathryn Muratore]] 10:20, 20 June 2011 (EDT)''':
** In retrospect, I'm not sure that I even used the correct tube here *'''[[User:Kathryn Muratore|Kathryn Muratore]] 16:26, 20 June 2011 (EDT)''':
*→ store @ -20°C
*→ store @ -20°C
 
==Results==
==Results==
{{#widget:Google Spreadsheet
{{#widget:Google Spreadsheet

Revision as of 13:26, 20 June 2011

AU CHEM-570 Lab Prep <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Bench work

  1. Miniprep O/N cultures from yesterday
  2. Quantify [minipreps]
    1. 95μL water + 5μL DNA from miniprep
      • → read A260 and A280
  3. NheI digest
    • performing sequential double digest, so use optimal buffer for SapI instead of NEB buffer 2
    • modify NheI digest protocol as follows
Tube sterile H2O 10X NEB4 100X BSA DNA NheI (10 kU/mL)
pTXB1His#1 34.5 μL 5 μL 0.5 μL 5 μL of 102 μg/mL 5μL
pTXB1 (82μg/mL) 34.5 μL 5 μL 0.5 μL 5 μL of 82 μg/mL 5μL
BSA PCR 24.5 μL 5 μL 0.5 μL 15 μL directly from PCR reaction 5μL
  • → 2h @ 37°C
  • → 20' @ 65°C (it took awhile to get the block to the right temp, so they were on ice for a bit, then at ~70°C before the block reached 65°C)
  • Note: I should have done a PCR cleanup on the BSA PCR sample! I will repeat this. *Kathryn Muratore 10:20, 20 June 2011 (EDT):
    • In retrospect, I'm not sure that I even used the correct tube here *Kathryn Muratore 16:26, 20 June 2011 (EDT):
  • → store @ -20°C

Results

{{#widget:Google Spreadsheet

key=0Aph9WCMwmkfMdF9PNG53aGllblpSLW1zdmpqcVR0TlE

}}

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