Difference between revisions of "Part:BBa K2889001:Design"
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https://static.igem.org/mediawiki/parts/9/91/Verify_pSB1C3-IL7-AS_by_sequencing.jpeg | https://static.igem.org/mediawiki/parts/9/91/Verify_pSB1C3-IL7-AS_by_sequencing.jpeg | ||
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===Source=== | ===Source=== | ||
Latest revision as of 08:23, 11 October 2018
pSB1C3-IL7-AS
- 10INCOMPATIBLE WITH RFC[10]Unknown
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23INCOMPATIBLE WITH RFC[23]Unknown
- 25INCOMPATIBLE WITH RFC[25]Unknown
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 127
Illegal BsaI.rc site found at 1183
Illegal SapI.rc site found at 429
Design Notes
IL7-AS [located anti-sense to interleukin-7 (Il7) gene; Accession number: NM_000880.3] is a newly discovered lncRNA that can be induced across multiple human and mouse cell types and has been reported to regulate the expression of interleukin-6 (IL6). We want to investigate the funtion of IL7-AS in kidney renal clear cell carcinoma. We cloned the full length of IL7-AS (3079bp) and inserted into pSB1C3 for submitting to IGEM 2018. We also inserted the full length of IL7-AS into pCDNA3.1 to study it's function in kidney renal clear cell carcinoma associated cell lines.
1.1 Amplification of IL7-AS fragments in human cells.
First we amplified IL7-AS fragment(line 2) 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 fragments to pSB1C3 vector. 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 full length of IL-AS from RNA of human cells.