Difference between revisions of "Part:BBa K2889002:Design"
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Latest revision as of 10:55, 11 October 2018
pSB1C3-IL7-AS-S1
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 600
Illegal SpeI site found at 813
Illegal SpeI site found at 924 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 626
Illegal SpeI site found at 813
Illegal SpeI site found at 924 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 600
Illegal SpeI site found at 813
Illegal SpeI site found at 924 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 600
Illegal SpeI site found at 813
Illegal SpeI site found at 924 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
We wanted to investigate which domain is essential for the function of IL7-AS. We cloned the truncated sequences of IL7-AS (IL7-AS-S1) into pSB1C3 for submitting to IGEM 2018 and inserted IL7-AS-S1 into pCDNA3.1 to study the function.
1.1 Amplification of IL7-AS-S1 fragments. First we amplified IL7-AS-S1 fragment (line 3) using primers. After that, we purified the PCR products by PCR Purification Kit and digested them with EcoRI and PstI (Fig 1).
1.2 Digested pSB1C3 vector.
We digested the pSB1C3 vectors with EcoRI and PstI (Fig 2).
1.3 Ligation of purified IL7-AS-S2; IL7-AS-S1 and IL7-AS fragments to PSB1C3 vector.
IL7-AS-S2; IL7-AS-S1 and IL7-AS fragments were ligated to PSB1C3 vector, respectively. Then we selected the positive clones by PCR and sequencing (Fig 3).
Source
We cloned the truncated sequences of IL7-AS (IL7-AS-S1, 1099 bp) from human cells