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← Primer database

Primer database

Acsl6

acyl-CoA synthetase long-chain family member 6

Mus musculus

PublicationZhang et al. 2022

Sequences

Forward (5′→3′)

AAGCAGTCGGAAGAAGTGGAG

Reverse (5′→3′)

GGCATCGTCATAGTAATGGGTAAG

Amplicon

93 bp

estimated from mapping

Primer Tm (°C)

ConditionsForwardReverseAverageΔ F/R
qPCR master mix63.562.062.71.5
Primer-BLAST default60.059.159.50.9

qPCR master mix: Owczarzy 2004/2008 · 250 nM primer · 50 mM Na⁺ · 3 mM Mg²⁺ · 0.8 mM dNTP

Primer-BLAST default: SantaLucia 1998 salt corr. · 50 nM primer · 50 mM Na⁺ · 1.5 mM Mg²⁺ · 0.6 mM dNTP

Bold, underlined base = 3′ end (polymerase extension point).

Primer QC

GC% · Forward

52.4%

GC% · Reverse

45.8%

Self-dimer ΔG · Fwd (any position)

-0.3 kcal/mol

Self-dimer ΔG · Fwd (3′ end)

No duplex found

Self-dimer ΔG · Rev (any position)

-0.3 kcal/mol

Self-dimer ΔG · Rev (3′ end)

No duplex found

Heterodimer ΔG (F×R, any position)

-0.3 kcal/mol

Heterodimer ΔG (F×R, 3′ end)

No duplex found

ΔG at 37 °C · SantaLucia 1998 NN parameters · < −6 kcal/mol = significant dimer risk · "3′ end" is restricted to duplexes anchored at a primer's own 3′ terminal base (polymerase-extendable); "any position" is the most stable duplex anywhere in the primer. Estimated screening value. For comprehensive dimer analysis, validate with Primer3.

Source

PMID

36438927(Europe PMC)

Citation

Zhang et al. 2022

Genomic mapping

Ensembl gene

ENSMUSG00000020333 ↗

NCBI Gene ID

216739

Assembly

GRCm39

Cross-check

✓ENSMUST00000093106
ENSMUST00000000145
✓ENSMUST00000138515
✓ENSMUST00000108904
✓ENSMUST00000108905
✓ENSMUST00000149403
✓ENSMUST00000072178
✓ENSMUST00000101211
✓ENSMUST00000101213★
✓ENSMUST00000064690
✓ENSMUST00000108899
✓ENSMUST00000094194
ENSMUST00000156252
ENSMUST00000139484
✓ENSMUST00000536508
✓ENSMUST00000536509
✓ENSMUST00000536510
✓ENSMUST00000536511
✓ENSMUST00000536513
✓ENSMUST00000536515
✓ENSMUST00000536517
✓ENSMUST00000536518
✓ENSMUST00000536519
✓ENSMUST00000536522
✓ENSMUST00000536523
✓ENSMUST00000536524
✓ENSMUST00000536525
✓ENSMUST00000536528
✓ENSMUST00000536529
✓ENSMUST00000536530
✓ENSMUST00000536531
✓ENSMUST00000536534
✓ENSMUST00000536535
✓ENSMUST00000536536
✓ENSMUST00000536538
✓ENSMUST00000536540
✓ENSMUST00000536541
✓ENSMUST00000536542
✓ENSMUST00000536543
✓ENSMUST00000536545
✓ENSMUST00000536546
✓ENSMUST00000536547
✓ENSMUST00000536548
✓ENSMUST00000536549
✓ENSMUST00000536550
✓ENSMUST00000536551
✓ENSMUST00000536552
✓ENSMUST00000536553
✓ENSMUST00000536554
✓ENSMUST00000536556
✓ENSMUST00000536557
✓ENSMUST00000536558
✓ENSMUST00000536559
✓ENSMUST00000536562
✓ENSMUST00000536563
✓ENSMUST00000536564
✓ENSMUST00000536565
✓ENSMUST00000536566
✓ENSMUST00000536567
✓ENSMUST00000536568
✓ENSMUST00000536569
✓ENSMUST00000536570
✓ENSMUST00000536571
✓ENSMUST00000536572
✓ENSMUST00000536573
✓ENSMUST00000536574
✓ENSMUST00000536575
✓ENSMUST00000536576
✓ENSMUST00000536577
✓ENSMUST00000536578
✓ENSMUST00000577758
✓ENSMUST00000577760
✓ENSMUST00000577761
96bp160bp~552bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.9.3kb65bp37bp32bp953bp~999bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.4.1kb130bp166bp~221bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.2.9kb115bp634bp65bp1.0kb~102bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.393bp100bp1.5kb179bp1.1kb~33bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.1.5kb52bp5.1kb74bp1.8kb~78bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.57bp~660bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.681bp~135bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.453bp~290bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.1.2kb96bp1.2kb73bp2.0kb127bp1.1kb~190bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.613bp~117bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.4.6kb~144bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.1.3kb102bp867bp~72bpThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.8.0kb~3.7kbThis is the outer span across every isoform's exon here. They don't all agree on the exact boundary (alternative splice site), so hover a row's own box below for its real length.
exon presentprimer binds hereconfirmed amplicongap = exon skipped~bp = boundary varies
How do I read this diagram?

Each row is one transcript, aligned on a shared axis. A gap means that isoform skips this exonic region. Each row's box is drawn at that isoform's own real exon length, which can be narrower than the column (alternative splice site); hover a box for its exact size. A ~amber header bp means isoforms don't all agree on this region's boundary, while a plain header means they do. Navy blocks mark where the primers bind, and green (with a ✓ by the name) means this isoform has a confirmed working amplicon, not just exon overlap. A row with a “‹ exons” / “exons ›” hint instead of any box means that isoform's exons exist but sit outside the current scroll position (a truncated isoform or alternate transcription start site) — click the hint to jump to them. Click a row to load it below.

★ = Ensembl canonical transcript (reference model, not necessarily the most expressed isoform)

Showing: ENSMUST00000093106

125bp116.2kb221bp22.9kb115bp3634bp65bp41.0kb102bp5393bp100bp61.5kb179bp71.1kb33bp81.5kb52bp95.1kb74bp101.8kb78bp111.4kb135bp12453bp135bp131.4kb96bp141.2kb73bp153.3kb89bp16613bp117bp174.6kb144bp181.3kb102bp19867bp72bp208.0kb3.7kb21FRPredicted from the amplicon's own GC%+length (von Ahsen et al. 2001, at the same qPCR buffer as the primer Tm above: 50 mM Na⁺ plus 3 mM Mg²⁺ less 0.8 mM dNTP, as a sodium equivalent), not the SantaLucia model used for primer Tm, which isn't valid at this length. A screening estimate for ranking amplicons — that relation was fitted on oligonucleotide duplexes, so applying it at amplicon length carries more error than its published ±1.9 °C, and it is not a melt-peak prediction.93 bp amplicon · predicted amplicon Tm 86.4 °C
Acsl6 · ENSMUST00000093106✓ Exon-junction spanning · gDNA-safe (>1.0kb intron)

Off-target specificity

Specific

No paired forward+reverse off-target amplicon found against nt (whole genome/transcriptome). Screen requires >=2 total mismatches including >=2 within the last 5bp at the 3' end to clear a hit (NCBI Primer-BLAST-style stringency); hits with >=6 total mismatches are dropped as implausible.

Efficiency measurements

Ideal range:Dilution series 1.90–2.10·LinRegPCR 1.70–1.90·unknown method: not scored
No measurements yet for this primer pair.

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