Difference between revisions of "Team:Aachen/Notebook/Documentation/Methanol Monocistronic Diversity Library"

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=Laboratory Notebook=
 
=Laboratory Notebook=
  
 +
===Constructing Precursor Plasmids===
 +
{|class="wikitable"
 +
! gene !! RDP parts !! product in !! expected length / result !! plasmid ID !! cryo ID
 +
|-
 +
| Mdh || (MDH.TER.AP00) #XOXM# + #PRDW# + #PLTB# + #PQT6# + #PVFL# || #K3YV# ; empty|| 3497 bp / crazy shit|| #CDPA#, #ZYBC# || #SF63#, #MBRR#
 +
|-
 +
| Mdh || (MDH.TER.AP04) #XOXM# + #PRDW# + #PLTB# + #WPW6# + #PVFL# || #AVQ6#; empty || 3497 bp / crazy shit || #YNOB#, #1KSW# || #DCN9#, #WHBF#
 +
|-
 +
| Mdh || (MDH.TER.AP10) #XOXM# + #PRDW# + #PLTB# + #3S8V# + #PVFL# || #ZE1A#; empty || 3497 bp / crazy shit || #EBWO#, #936C# || #8KAA#, #EB88#
 +
|-
 +
| Mdh || (MDH.TER.AP19) #XOXM# + #PRDW# + #PLTB# + #MBWS# + #PVFL# || #NDKS#; empty  || 3497 bp / crazy shit || #1MC6#, #ESQD# || #48C8#, #8BZ9#
 +
|}
  
 +
*heat shocked in DH5alpha with 5 µl of DNA
 +
 +
==15-08-09==
 +
 +
'''Gel control of MDH precursor plasmids''':
 +
*no good picture could be taken because the UV light destroyed the DNA; picture of it will be taken tomorrow in the Schwaneberg lab
 +
*master plates and overnights of the backup plasmids
 +
*plasmid prep and cryos of MDH precursor plasmids
 +
 +
==15-08-10==
 +
*make cryos and prepp plasmids of MDH precursor plasmids
 +
*Measure concentrations of prepped MDH precursor plasmids
 +
*Gel control of MDH precursor plasmids in Schwaneberg Lab
 +
*Measure concentrations of prepped backup plasmids
 +
*Pipett sequencing of backup plasmids and MDH precursor plasmids
 +
 +
* test digest of backup and MDH precursor plasmids:
 +
 +
{|class="wikitable"
 +
! digest ID !! template ID !! expected lengths [bp] !! result
 +
|-
 +
| A || #CDPA# || 2226 / 1279 || 2226 / -
 +
|-
 +
| B || #ZYBC# || 2226 / 1279 || 2226 / 1700
 +
|-
 +
| C || #YNOB# || 2226 / 1279 || 2226 / 1700
 +
|-
 +
| D || #1KSW# || 2226 / 1279 || 2226 / 1700
 +
|-
 +
| E || #EBWO# || 2226 / 1279 || 2226 / -
 +
|-
 +
| F || #936C# || 2226 / 1279 || 2226 / -
 +
|-
 +
| G || #1MC6# || 2226 / 1279 || 2226 / 1279
 +
|-
 +
| H || #ESQD# || 2226 / 1279 || 2226 / 1279
 +
|-
 +
| I || #LWDR# || 2226 / 1187 || 2226 / 1187 + one long band
 +
|-
 +
| J || #4TSQ# || 2226 / 1187 || 2226 / 1187
 +
|-
 +
| K || #KTHK# || 2226 / 665 || 2226 / 665
 +
|-
 +
| L || #O9ZE# || 2226 / 584 || 2226 / 584
 +
|-
 +
| M || #6QE3# || 2226 / 2644 || 2226 / 2644 + one short band
 +
|-
 +
| N || #KVEY# || 2226 / 2644 || 2226 / 2644 + two long bands
 +
|}
 +
 +
digests stored in falcon #PSHF#
  
 
=References=
 
=References=

Revision as of 10:44, 18 September 2015


Laboratory Notebook

Constructing Precursor Plasmids

gene RDP parts product in expected length / result plasmid ID cryo ID
Mdh (MDH.TER.AP00) #XOXM# + #PRDW# + #PLTB# + #PQT6# + #PVFL# #K3YV# ; empty 3497 bp / crazy shit #CDPA#, #ZYBC# #SF63#, #MBRR#
Mdh (MDH.TER.AP04) #XOXM# + #PRDW# + #PLTB# + #WPW6# + #PVFL# #AVQ6#; empty 3497 bp / crazy shit #YNOB#, #1KSW# #DCN9#, #WHBF#
Mdh (MDH.TER.AP10) #XOXM# + #PRDW# + #PLTB# + #3S8V# + #PVFL# #ZE1A#; empty 3497 bp / crazy shit #EBWO#, #936C# #8KAA#, #EB88#
Mdh (MDH.TER.AP19) #XOXM# + #PRDW# + #PLTB# + #MBWS# + #PVFL# #NDKS#; empty 3497 bp / crazy shit #1MC6#, #ESQD# #48C8#, #8BZ9#
  • heat shocked in DH5alpha with 5 µl of DNA

15-08-09

Gel control of MDH precursor plasmids:

  • no good picture could be taken because the UV light destroyed the DNA; picture of it will be taken tomorrow in the Schwaneberg lab
  • master plates and overnights of the backup plasmids
  • plasmid prep and cryos of MDH precursor plasmids

15-08-10

  • make cryos and prepp plasmids of MDH precursor plasmids
  • Measure concentrations of prepped MDH precursor plasmids
  • Gel control of MDH precursor plasmids in Schwaneberg Lab
  • Measure concentrations of prepped backup plasmids
  • Pipett sequencing of backup plasmids and MDH precursor plasmids
  • test digest of backup and MDH precursor plasmids:
digest ID template ID expected lengths [bp] result
A #CDPA# 2226 / 1279 2226 / -
B #ZYBC# 2226 / 1279 2226 / 1700
C #YNOB# 2226 / 1279 2226 / 1700
D #1KSW# 2226 / 1279 2226 / 1700
E #EBWO# 2226 / 1279 2226 / -
F #936C# 2226 / 1279 2226 / -
G #1MC6# 2226 / 1279 2226 / 1279
H #ESQD# 2226 / 1279 2226 / 1279
I #LWDR# 2226 / 1187 2226 / 1187 + one long band
J #4TSQ# 2226 / 1187 2226 / 1187
K #KTHK# 2226 / 665 2226 / 665
L #O9ZE# 2226 / 584 2226 / 584
M #6QE3# 2226 / 2644 2226 / 2644 + one short band
N #KVEY# 2226 / 2644 2226 / 2644 + two long bands

digests stored in falcon #PSHF#

References

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