Revathireddy (Talk | contribs) |
Revathireddy (Talk | contribs) |
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<h2>Week 20: On our toes!</h2> | <h2>Week 20: On our toes!</h2> | ||
− | <p style="font-family: 'Lato'; font-size:15px">We immediately designed and ordered primers to amplify the ychH promoter from | + | <p style="font-family: 'Lato'; font-size:15px">We immediately designed and ordered primers to amplify the <i>ychH</i> promoter from <i>E. coli</i> MG1655 genomic DNA. The ychH promoter was inoculated in LB+Kanamycin and miniprepped the next day.We await our primers!<br><br> |
Meanwhile, ligated products (<i>glnA</i> + T7 + lacI with pSB3C5) were transformed in <i>E. coli</i> DH5-α. Three colonies were picked up randomly and inoculated in LB+chloramphenicol. Along with that, <i>E. coli</i> DH5-α transformed with BBa_K1152013 was also inoculated in LB+chloramphenicol.Both were miniprepped the next day.Both were double digested with EcoRI and PstI and a ligation reaction was setup in 1:3 molar ratio. This ligated product was transformed in <i>E. coli</i> DH5-α. Many red colonies were seen the next day which indicate self-ligation. Other colonies were screened and found to be negative | Meanwhile, ligated products (<i>glnA</i> + T7 + lacI with pSB3C5) were transformed in <i>E. coli</i> DH5-α. Three colonies were picked up randomly and inoculated in LB+chloramphenicol. Along with that, <i>E. coli</i> DH5-α transformed with BBa_K1152013 was also inoculated in LB+chloramphenicol.Both were miniprepped the next day.Both were double digested with EcoRI and PstI and a ligation reaction was setup in 1:3 molar ratio. This ligated product was transformed in <i>E. coli</i> DH5-α. Many red colonies were seen the next day which indicate self-ligation. Other colonies were screened and found to be negative |
Revision as of 07:37, 1 November 2017