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← Primer database

Primer database

METTL14

methyltransferase 14, N6-adenosine-methyltransferase non-catalytic subunit

Homo sapiens

PublicationChen et al. 2024

Sequences

Forward (5′→3′)

AGAGAACAAAGGAACACTGCCT

Reverse (5′→3′)

AATGAAGTCCCCGTCTGTGC

Amplicon

71 bp

estimated from mapping

Primer Tm (°C)

ConditionsForwardReverseAverageΔ F/R
qPCR master mix63.164.063.60.9
Primer-BLAST default59.860.360.10.5

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

45.5%

GC% · Reverse

55.0%

Self-dimer ΔG · Fwd (any position)

-1.1 kcal/mol

Self-dimer ΔG · Fwd (3′ end)

-1.1 kcal/mol

Self-dimer ΔG · Rev (any position)

-0.3 kcal/mol

Self-dimer ΔG · Rev (3′ end)

-0.3 kcal/mol

Heterodimer ΔG (F×R, any position)

-1.1 kcal/mol

Heterodimer ΔG (F×R, 3′ end)

-0.3 kcal/mol

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

39720723(Europe PMC)

Citation

Chen et al. 2024

Genomic mapping

Ensembl gene

ENSG00000145388 ↗

NCBI Gene ID

57721

Assembly

GRCh38

Cross-check

✓ENST00000388822★
ENST00000508801
ENST00000506780
✓ENST00000900246
✓ENST00000900247
✓ENST00000900248
ENST00000900250
✓ENST00000949682
ENST00000975295
ENST00000975296
ENST00001089502
ENST00001105516
✓ENST00001140038
~220bpThis 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.949bp150bp1.2kb89bp1.4kb~88bpThis 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.2.1kb81bp368bp~97bpThis 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.2.4kb~91bpThis 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.2.7kb~142bpThis 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.3.2kb~952bpThis 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.441bp50bp893bp50bp999bp~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.1.6kb~211bpThis 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.4.2kb~5.4kbThis 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: ENST00000388822 ★

209bp12.3kb89bp21.4kb88bp32.1kb81bp4368bp88bp52.4kb91bp62.7kb142bp73.2kb93bp83.3kb117bp91.6kb211bp104.2kb5.4kb11FRPredicted 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.71 bp amplicon · predicted amplicon Tm 82.4 °C
METTL14 · ENST00000388822 (NM_020961) · canonical✓ Exon-junction spanning · gDNA-safe by primer design

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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