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← Primer database

Primer database

GUSB

glucuronidase beta

Homo sapiens

PublicationDong et al. 2022⇄ Compare with 1 other candidate →

Sequences

Forward (5′→3′)

AGCCAGTTCCTCATCAATGG

Reverse (5′→3′)

GGTAGTGGCTGGTACGGAAA

Amplicon

160 bp

Primer Tm (°C)

ConditionsForwardReverseAverageΔ F/R
qPCR master mix61.363.262.21.9
Primer-BLAST default57.659.458.51.8

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

50.0%

GC% · Reverse

55.0%

Self-dimer ΔG · Fwd (any position)

-1.3 kcal/mol

Self-dimer ΔG · Fwd (3′ end)

-1.3 kcal/mol

Self-dimer ΔG · Rev (any position)

-1.6 kcal/mol

Self-dimer ΔG · Rev (3′ end)

No duplex found

Heterodimer ΔG (F×R, any position)

-4.8 kcal/mol

Heterodimer ΔG (F×R, 3′ end)

1.0 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

36568365(Europe PMC)

Citation

Dong et al. 2022

Genomic mapping

Ensembl gene

ENSG00000169919 ↗

NCBI Gene ID

2990

Assembly

GRCh38

Cross-check

✓ENST00000304895★
✓ENST00000421103
ENST00000864776
ENST00000864777
✓ENST00000864778
ENST00000864779
✓ENST00000864780
✓ENST00000864781
ENST00000864782
✓ENST00000864783
ENST00000864784
✓ENST00000864785
✓ENST00000864786
ENST00000864787
✓ENST00000864788
ENST00000864789
✓ENST00000864790
ENST00000864791
✓ENST00000864792
✓ENST00000864793
ENST00000864794
ENST00000864795
✓ENST00000864796
ENST00000864797
✓ENST00000864798
ENST00000912571
ENST00000912572
ENST00000912573
ENST00000912574
ENST00000912575
ENST00000912576
✓ENST00000912577
ENST00000912578
ENST00000952458
ENST00000952459
ENST00000952460
ENST00000952461
✓ENST00000986743
ENST00000986748
ENST00001100603
ENST00001100605
ENST00001100606
ENST00001100613
ENST00001100614
✓ENST00001100619
ENST00001100623
✓ENST00001100629
ENST00001100632
✓ENST00001100634
✓ENST00001100636
ENST00001100640
✓ENST00001142510
~310bpThis 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.~270bpThis 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.~248bpThis 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.~170bpThis 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.~219bpThis 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.~153bpThis 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.179bp~147bpThis 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.~85bpThis 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.~177bpThis 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.~136bpThis 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.21bp~387bpThis 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: ENST00000304895 ★

240bp12.0kb186bp2681bp185bp3511bp143bp43.5kb188bp51.3kb153bp6544bp179bp7408bp147bp84.2kb85bp92.6kb177bp103.6kb136bp113.8kb380bp12FRPredicted 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.160 bp amplicon · predicted amplicon Tm 89.1 °C
GUSB · ENST00000304895 (NM_000181) · canonical⚠ Exon-junction spanning · gDNA product still possible (300bp–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.

Mean efficiency · Standard curve

1.970

from 1 measurement

Efficiency measurements

Ideal range:Dilution series 1.90–2.10·LinRegPCR 1.70–1.90·unknown method: not scored
EfficiencyR²MethodSourcePCR mixInstrumentAnnealingn
1.97097.0%
—Literature-reportedvia dilution seriesPMID 36568365All-in-One qPCR Mix (GeneCopoeia)Roche LightCycler 480 (Roche Diagnostics, Germany)55 °C3

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