Difference between revisions of "Part:BBa K2719005:Experience"
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[[file:T--TecCEM--ResK2719005.png|500px]] | [[file:T--TecCEM--ResK2719005.png|500px]] | ||
| − | <p><i>Figure 3.</i> 0.85% agarose gel with GelRed: MW) | + | <p><i>Figure 3.</i> 0.85% agarose gel with GelRed: MW) 2-Log DNA Ladder; 4) Tenascin 5 Domain V Expression device (BBa_K2719005) restriction with NotI</p> |
<p>The <i>E.coli</i> BL21 (DE3) colonies were later inoculated on liquid media (seed culture) and incubated overnight. From the seed culture 1 ml was inoculated on fresh LB+CAM broth, and at regular intervals the absorbance was measured. When the absorbance reached 0.7, IPTG was added to a final concentration of 1 mM and samples were taken after 5 and 16 h. Those samples were lysed. The lysate was loaded into a 18% polyacrylamide SDS-PAGE (Figure 4), from which a higher presence of protein was detected at 5 h after induction.</p> | <p>The <i>E.coli</i> BL21 (DE3) colonies were later inoculated on liquid media (seed culture) and incubated overnight. From the seed culture 1 ml was inoculated on fresh LB+CAM broth, and at regular intervals the absorbance was measured. When the absorbance reached 0.7, IPTG was added to a final concentration of 1 mM and samples were taken after 5 and 16 h. Those samples were lysed. The lysate was loaded into a 18% polyacrylamide SDS-PAGE (Figure 4), from which a higher presence of protein was detected at 5 h after induction.</p> | ||
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| + | [[file:T--TecCEM--SDSK2719005.png|500px]] | ||
| + | <p><i>Figure 4.</i> 20% polyacrylamide gel: MW) Presicion Plus Protein Standard; 7) Tenascin 5 Domain V Expression device (BBa_K2719005) protein</p> | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 22:54, 17 October 2018
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how you used this part and how it worked out.
Applications of BBa_K2719005
This part was cloned on pSB1C3 using EcoRI and PstI, later it was transformed on E.coli DH5a (Figure 1).
Figure 1. Colonies transformed with the Tenascin 5 Domain V Expression device
Then the plasmid was extracted and run on an agarose gel electrophoresis (Figure 2). Latter an restriction was performed using NotI and was run on an agarose gel (Figure 3) to probe the presence of the insert. After the extracted plasmid it was transformed on E.coli BL21 (DE3).
Figure 2. 0.85% agarose gel with GelRed: MW) 1Kb plus from NEB; 7) Tenascin 5 Domain V Expression device (BBa_K2719005) V'; 8) Tenascin 5 Domain V Expression device (BBa_K2719005) V
Figure 3. 0.85% agarose gel with GelRed: MW) 2-Log DNA Ladder; 4) Tenascin 5 Domain V Expression device (BBa_K2719005) restriction with NotI
The E.coli BL21 (DE3) colonies were later inoculated on liquid media (seed culture) and incubated overnight. From the seed culture 1 ml was inoculated on fresh LB+CAM broth, and at regular intervals the absorbance was measured. When the absorbance reached 0.7, IPTG was added to a final concentration of 1 mM and samples were taken after 5 and 16 h. Those samples were lysed. The lysate was loaded into a 18% polyacrylamide SDS-PAGE (Figure 4), from which a higher presence of protein was detected at 5 h after induction.
Figure 4. 20% polyacrylamide gel: MW) Presicion Plus Protein Standard; 7) Tenascin 5 Domain V Expression device (BBa_K2719005) protein
User Reviews
UNIQdb3aa87048ed2dc6-partinfo-00000000-QINU UNIQdb3aa87048ed2dc6-partinfo-00000001-QINU