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Team:Freiburg Bioware/NoteBook/Labjournal/October
From 2010.igem.org
(Difference between revisions)
(→136. labday 01.10.2010) |
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'''Investigator: Kira''' | '''Investigator: Kira''' | ||
| - | c(pSB1C3_lITR_hTERT_beta-globin)= 333 ng/ul | + | c(pSB1C3_lITR_hTERT_beta-globin)= 333 ng/ul <br /> |
| - | c(pSB1C3_CD)= 151 ng/ul | + | c(pSB1C3_CD)= 151 ng/ul <br /> |
| - | | border="1" | + | {| border="1" |
| align="left" | '''Components''' ||align="left"| <b>vector</b> Volume/µL ||align="left"| <b>insert</b> Volume/µL | | align="left" | '''Components''' ||align="left"| <b>vector</b> Volume/µL ||align="left"| <b>insert</b> Volume/µL | ||
|- | |- | ||
| Line 67: | Line 67: | ||
1% agarose gel <br /> | 1% agarose gel <br /> | ||
| + | |||
| + | [[Image: Freiburg10_CD-beta-globin.jpg]] | ||
| + | |||
| + | '''Ligation'''<br /> | ||
| + | |||
| + | DNA-mix: 8 ul (vector..)+(insert...) <br /> | ||
| + | T4 ligase: 1 ul <br /> | ||
| + | T4 buffer: 1 ul <br /> | ||
| + | Incubation @ RT for 30 min <br /> | ||
| + | |||
| + | Transformation was performed according to the standard protocol w BL21 cells. <br /> | ||
Revision as of 17:27, 1 October 2010
Contents |
136. labday 01.10.2010
mini preps of CD clones
Investigator: Kira
c(p689)= 115 ng/ul
c(690)= 151, 20 ng/ul
c(691)= 122,27 ng/ul
c(p692) = 126,42 ng/ul
test digestion of CD clones
Investigator: Kira
| Components | sample Volume/µL |
| DNA | 4,0 µl |
| BSA (10x) | 2 µl |
| Buffer no. 4 | 2,0 µl |
| Enzyme 1 XbaI | 1,0 µl |
| Enzyme 2 AgeI | 1,5 µl |
| H2O | 9,5 µl |
| Total volume | 20 |
incubation @ 37 C for approx. 2 h
1% agarose gel
Biobrick assembly pSB1C3_lITR_hTERT_beta-globin_CD
Investigator: Kira
c(pSB1C3_lITR_hTERT_beta-globin)= 333 ng/ul
c(pSB1C3_CD)= 151 ng/ul
| Components | vector Volume/µL | insert Volume/µL |
| DNA | 4,5 µl | 6 |
| BSA (10x) | 3 µl | 3 |
| Buffer no. 4 | 3,0 µl | 3 |
| Enzyme 1 XbaI | 0 µl | 1,5 |
| Enzyme 2 SpeI | 1,5 µl | 0 |
| Enzyme 3 PstI-HF | 1,0 | 1 |
| H2O | 17 | 15,5 |
| Total volume | 25 |
incubation @ 37 C for approx. 2 h
1% agarose gel
Ligation
DNA-mix: 8 ul (vector..)+(insert...)
T4 ligase: 1 ul
T4 buffer: 1 ul
Incubation @ RT for 30 min
Transformation was performed according to the standard protocol w BL21 cells.
