Imperial/Wet Lab/Lab Notebook/2007-09-14
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+ | ==Restriction Digest of Biobricks== | ||
+ | #4 DNA samples were digested by SpeI and PstI | ||
+ | #* Samples 5, 7, 24 | ||
+ | #1 DNA sample wasdigested by XbaI and PstI | ||
+ | #* Samples 23 | ||
+ | #4 DNA samples were digested by EcoRI | ||
+ | #* Samples 13, 25, 27b-e, T7, Kirsten's parts | ||
+ | #4 μl of DNA was digested by 1 μl of enzymes at 37°C for 1 hour<br>Exception: Sample 23 had 6 μl DNA and 2 μl enzymes | ||
+ | |||
+ | Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Restriction_Digest | Restriction Digest] in the general protocols page | ||
+ | |||
+ | ==Agarose Gel Electrophoresis of Biobricks== | ||
+ | #Checked 9 Biobricks and 13 Digests on 1% agarose gel | ||
+ | [[Image:ICGEMS Gel 14 9.png|900px|]] | ||
+ | |||
+ | Conclusions: | ||
+ | *Part 27 is confirmed to be faulty | ||
+ | *T7 maxi-prep was not done properly | ||
+ | *Kirsten's parts were most likely not there... | ||
+ | |||
+ | Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Electrophoresis | Electrophoresis] in the general protocols page | ||
+ | |||
+ | ==Gel Purification of Biobricks== | ||
+ | |||
+ | #Gel was melted and run through column | ||
+ | #Purified DNA was eluted with ddH<sub>2</sub>O | ||
+ | <font size=-2> | ||
+ | *5. BBa_R0040 [ptet] (~2100 bp fragment) | ||
+ | *7. BBa_F2620 [ptet-LuxR-pLux] (~3100 bp fragment) | ||
+ | *23. BBa_I13504 [GFP] (~800 bp fragment) | ||
+ | *24. BBa_R0062 [pLux] (~2100 bp fragment) | ||
+ | </font> | ||
+ | |||
+ | Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Gel_Purification | Gel Purification] in the general protocols page | ||
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Latest revision as of 19:01, 26 October 2007
14 September 2007
Restriction Digest of Biobricks
- 4 DNA samples were digested by SpeI and PstI
- Samples 5, 7, 24
- 1 DNA sample wasdigested by XbaI and PstI
- Samples 23
- 4 DNA samples were digested by EcoRI
- Samples 13, 25, 27b-e, T7, Kirsten's parts
- 4 μl of DNA was digested by 1 μl of enzymes at 37°C for 1 hour
Exception: Sample 23 had 6 μl DNA and 2 μl enzymes
Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Restriction_Digest | Restriction Digest] in the general protocols page
Agarose Gel Electrophoresis of Biobricks
- Checked 9 Biobricks and 13 Digests on 1% agarose gel
Conclusions:
- Part 27 is confirmed to be faulty
- T7 maxi-prep was not done properly
- Kirsten's parts were most likely not there...
Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Electrophoresis | Electrophoresis] in the general protocols page
Gel Purification of Biobricks
- Gel was melted and run through column
- Purified DNA was eluted with ddH2O
- 5. BBa_R0040 [ptet] (~2100 bp fragment)
- 7. BBa_F2620 [ptet-LuxR-pLux] (~3100 bp fragment)
- 23. BBa_I13504 [GFP] (~800 bp fragment)
- 24. BBa_R0062 [pLux] (~2100 bp fragment)
Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Gel_Purification | Gel Purification] in the general protocols page