Part:BBa_K4630103:Experience

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Applications of BBa_K4630103

We cultured the successfully induced stgRNA barcode cassette 1 monoclonal colony overnight on a 37 degree shaker to remove the pCas plasmid.We used SpeI and EcoRI to cut the stgRNA-barcode-cassette 1 plasmid. Then, XbaI and EcoRI were used to cut the recording fragments from the stgRNA-barcode-cassette 2 plasmid. Due to the ability of SpeI and XbaI to form the same viscous end, we were able to connect the carrier and fragment to each other by holding them at 37 degrees Celsius for 10 minutes using T4 ligase. Through sequencing, we successfully screened the successfully connected monoclonal colonies(Fig 1).

Figure. 1 Sequencing results of stgRNA barcode cassette (1+2) and stgRNA barcode cassette (1-1+2).

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