User:Karmella Haynes/Notebook/BioBrick cloning/2015/05/06: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 57: Line 57:
|-
|-
| 4. pBja-BjaR/MRV-2 || 0.043 || 1.934 || 43.1
| 4. pBja-BjaR/MRV-2 || 0.043 || 1.934 || 43.1
|-
| 5. pBra-BraR/MRV-1 || 0.070 || 2.021 || 69.9
|-
| 6. pBra-BraR/MRV-2 || 0.030 || 1.946 || 29.8
|-
| 7. pRpa-RpaR/MRV-1 || 0.093 || 1.956 || 92.6
|-
| 7. pRpa-RpaR/MRV-2 || 0.033 || 2.000 || 32.7
|}
|}


Revision as of 19:11, 6 May 2015

Karmella's BioBrick Cloning <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>

05/05/15

  • Ryan - minipreps & digests


Minipreps

  • Use Sigma GenElute; elute with 75 μL elution solution
  • promoter/Regulator/MRV colonies
  • Cut w/ KpnI/EcoRI - BbsI drop-in of promoter destroys KpnI site; empty vector will produce 2 bands, successful clones will be linearized (1 cut)
  1. pAub-AubR/MRV
  2. pBja-BjaR/MRV
  3. pBra-BraR/MRV
  4. pRpa-RpaR/MRV


Reagent Volume Expected:1,2. pAub/AubR/MRV = 4240
3,4. pBja/BjaR/MRV = 4099
5,6. pBra/BraR/MRV = ###
7,8. pRpa/RpaR/MRV = 4116
Empty vector = ~4000, 971 		Hover name
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 3.0 μL
10X buffer 1.5
EcoRI 1.0
KpnI 1.0
dH2O 8.5
  15 μL

--> 37°C/ ~10 min.


  • Measure [DNA]
Sample OD 260 260/280 ng/μL
1. pAub-AubR/MRV-1 0.152 1.930 152.3
2. pAub-AubR/MRV-2 0.079 1.992 78.6
3. pBja-BjaR/MRV-1 0.093 1.989 92.7
4. pBja-BjaR/MRV-2 0.043 1.934 43.1
5. pBra-BraR/MRV-1 0.070 2.021 69.9
6. pBra-BraR/MRV-2 0.030 1.946 29.8
7. pRpa-RpaR/MRV-1 0.093 1.956 92.6
7. pRpa-RpaR/MRV-2 0.033 2.000 32.7


Conclusions

  • tba



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Karmella_Haynes/Notebook/BioBrick_cloning/2015/05/06&oldid=883591"