Difference between revisions of "Team:UCLA/Notebook/Spider Silk Genetics/22 July 2015"
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*The digestion worked, but it is not as complete as previous digestions have been. Compare to [https://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/13_July_2015#BsaI_Digestion_of_MaSp2_Plasmids_and_Gel_Purification previously]. | *The digestion worked, but it is not as complete as previous digestions have been. Compare to [https://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/13_July_2015#BsaI_Digestion_of_MaSp2_Plasmids_and_Gel_Purification previously]. | ||
− | ==Midi prep for M2-AB | + | ==Midi prep for M2-AB== |
+ | *Used Qiagen Kit, eluted in 200 uL EB. | ||
+ | *Approximately 100 ng/uL for each) | ||
+ | *May need to only digest 4 ug of each in the future. | ||
+ | |||
+ | ==Bacterial Culture== | ||
+ | *Picked three colonies from each plate from [https://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/21_July_2015#Transformation yesterday]. | ||
+ | **Grow in 5 uL liquid culture. | ||
+ | *Streaked one plate with all three of the M2-9(T7) samples we sent for sequencing. |
Latest revision as of 04:01, 27 July 2015
Contents
7/22/2015
Transformation Results
- Colonies are present on all plates from yesterday.
Miniprep
- Used Zymo Classic kit to prep M2-9(T7) and M2-SeqAB
Concentration (ng/uL) | A 260/280 | |
---|---|---|
M2-9(T7)-1 | 590.49 | 1.86 |
M2-9(T7)-2 | 614.07 | 1.84 |
M2-9(T7)-3 | 392.19 | 1.84 |
M2-Seq AB | 1117.69 | 1.88 |
- Sent M2-9(T7) samples for sequencing using VF/R primers as needed.
BsaI Digestion for M2-Seq AB
- Digest 5 ug of M2-SeqAB plasmid in 50 uL reaction with 4 uL of BsaI.
Volume (uL) | |
---|---|
BsaI | 4 |
10x Cutsmart | 5 |
DNA | 4.47 |
ddH2O | 36.53 |
- Ran digest on 1.5% TAE gel. Used 2 uL of 50 bp ladder.
- The digestion worked, but it is not as complete as previous digestions have been. Compare to previously.
Midi prep for M2-AB
- Used Qiagen Kit, eluted in 200 uL EB.
- Approximately 100 ng/uL for each)
- May need to only digest 4 ug of each in the future.
Bacterial Culture
- Picked three colonies from each plate from yesterday.
- Grow in 5 uL liquid culture.
- Streaked one plate with all three of the M2-9(T7) samples we sent for sequencing.