Part:BBa_K4717000
EIF6 primer
Primer sequences are essential components in polymerase chain reaction (PCR) and similar molecular biology techniques. They are short, single-stranded DNA sequences designed to bind to specific regions of a target DNA molecule, serving as the starting point for DNA synthesis during PCR amplification. The design of primer sequences involves considerations such as length, GC content, melting temperature (Tm), and specificity, all of which impact the success of DNA amplification. In PCR applications, primer sequences play a crucial role in DNA amplification, gene expression analysis, genotyping, mutation detection, and many other genetic investigations, making them fundamental tools in molecular biology research. In this project, we used Primer3 (https://bioinfo.ut.ee/primer3-0.4.0/) to design the primer sequence based on information of NCBI nucleoside database. We designed three primers for each gene and selected one with the best specificity and efficiency. The designed primary was then synthesized by solid-phase oligonucleotide synthesis method using an automated DNA synthesizer.
the sequece of three primer was showing as following：
Primer 1：
| OLIGO | Start | Length | Tm | GC% | Any | 3' | Rep | Sequence |
|---|---|---|---|---|---|---|---|---|
| LEFT | 1267 | 20 | 59.94 | 55.00 | 3.00 | 1.00 | 10.00 | TCTCATCTCTCCCTGCCACT |
| RIGHT | 1510 | 20 | 60.00 | 50.00 | 3.00 | 0.00 | 10.00 | GCAACAAGAGCGAAAACTCC |
SEQUENCE SIZE: 5867 INCLUDED REGION SIZE: 5867 PRODUCT SIZE: 244, PAIR ANY COMPL: 4.00, PAIR 3' COMPL: 0.00
Primer 2：
| OLIGO | Start | Length | Tm | GC% | Any | 3' | Sequence |
|---|---|---|---|---|---|---|---|
| LEFT | 1267 | 20 | 59.94 | 55.00 | 3.00 | 1.00 | TCTCATCTCTCCCTGCCACT |
| RIGHT | 1511 | 20 | 60.00 | 50.00 | 3.00 | 0.00 | GGCAACAAGAGCGAAAACTC |
PRODUCT SIZE: 245, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00 Primer 3：
| OLIGO | Start | Length | Tm | GC% | Any | 3' | Sequence |
|---|---|---|---|---|---|---|---|
| LEFT | 4204 | 20 | 59.97 | 55.00 | 3.00 | 0.00 | AGGGGTCTCTCCTGTGGTTT |
| RIGHT | 4353 | 20 | 59.93 | 50.00 | 4.00 | 2.00 | GCAGGACTGGATGGTTCAAT |
SEQUENCE SIZE: 5867 PRODUCT SIZE: 191, PAIR ANY COMPL: 3.00, PAIR 3' COMPL: 1.00 Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| None |