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← Primer database

Primer database

ACTB

actin beta

Homo sapiens

PublicationXiao et al. 2022⇄ Compare with 3 other candidates →

Sequences

Forward (5′→3′)

GCCGACAGGATGCAGAAGG

Reverse (5′→3′)

TGGAAGGTGGACAGCGAGG

Amplicon

131 bp

estimated from mapping

Primer Tm (°C)

ConditionsForwardReverseAverageΔ F/R
qPCR master mix64.965.965.41.0
Primer-BLAST default60.861.661.20.8

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

63.2%

GC% · Reverse

63.2%

Self-dimer ΔG · Fwd (any position)

-3.1 kcal/mol

Self-dimer ΔG · Fwd (3′ end)

0.1 kcal/mol

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)

0.1 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

36514742(Europe PMC)

Citation

Xiao et al. 2022

Genomic mapping

Ensembl gene

ENSG00000075624 ↗

NCBI Gene ID

60

Assembly

GRCh38

Cross-check

✓ENST00000674681
✓ENST00000642480
✓ENST00000676397
ENST00000676319
✓ENST00000473257
✓ENST00000646664★
✓ENST00000464611
✓ENST00000675515
✓ENST00000493945
ENST00000432588
ENST00000645576
ENST00000647275
ENST00000443528
ENST00000417101
ENST00000414620
✓ENST00000899420
✓ENST00000899421
✓ENST00000899422
ENST00000899423
✓ENST00000899424
✓ENST00000899425
✓ENST00000899426
✓ENST00000899427
ENST00000899428
✓ENST00000899429
✓ENST00000919298
✓ENST00000919299
ENST00000919300
ENST00000919301
ENST00000919302
✓ENST00000919303
✓ENST00000919304
✓ENST00000919305
✓ENST00000919306
✓ENST00000950579
ENST00001073015
ENST00001073020
ENST00001099860
ENST00001099861
ENST00001099863
✓ENST00001099864
ENST00001099865
✓ENST00001099867
✓ENST00001141547
189bp~255bpThis 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.84bp96bp~879bpThis 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.82bp80bp~448bpThis 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.~240bpThis 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.~439bpThis 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.~1.8kbThis 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: ENST00000674681

60bp11.1kb150bp2522bp240bp31.1kb439bp4714bp182bp51.8kb1.5kb6FRPredicted 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.131 bp amplicon · predicted amplicon Tm 89.7 °C
ACTB · ENST00000674681✓ 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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