ExoSAP-IT PCR cleanup reagents

One step to superior sequencing results

ExoSAP-IT Express reagent: PCR cleanup Fastest PCR cleanup method
in as little as 5 minutes ExoSAP-IT Express reagent includes a novel exonuclease
I that removes unincorporated primers and nucleotides
Our one-step Applied Biosystems™ ExoSAP-IT™ Express with a reduced purification time. Only one pipetting step is
PCR Product Cleanup Reagent enables quality sequencing required, simplifying the workflow and producing a sample
results in a fraction of the time. ready for downstream applications in only 5 minutes
(Figure 2).
• 5 min protocol—fastest enzymatic cleanup of
PCR product

• One-tube, one-step PCR cleanup—add reagent Magnetic ExoSAP-IT Express Spin

beads vs. reagent vs. columns
directly to PCR product

• Novel enzyme technology—enzymes irreversibly Nine Eight

steps steps
inactivated in just 1 min at 80°C One
pipette
step
• Conserve PCR samples—100% recovery of PCR
products, regardless of amplicon length
45 min protocol 5 min protocol 20 min protocol

• Scalable—treat PCR reaction in volumes from 5 µL

to 5 mL Figure 2. Comparison of cleanup methods. Use of ExoSAP-IT Express
reagent eliminates spin columns, magnetic beads, filtration, and gel
• Eliminate spin columns or magnetic beads—helps purification. With a 5 min protocol, ExoSAP-IT Express reagent is the
fastest and easiest method for PCR cleanup, helping to minimize pipetting
decrease time and expense while increasing yield errors and contamination.

High-quality, accurate results Conserve PCR samples—simple one-step,

Compared with alternative PCR cleanup methods, 100% recovery
ExoSAP-IT Express PCR Product Cleanup Reagent helps The ExoSAP-IT Express enzymatic cleanup method
to ensure the availability of purified samples ready for helps minimize errors by reducing your protocol to a
downstream applications in just 5 minutes. This unique, single pipetting step, allowing for automated or manual
highly stable one-tube solution allows for 100% recovery processing. ExoSAP-IT Express reagent outperforms the
of DNA and longer read lengths for greater confidence, competition with 100% recovery of all amplicon sizes, from
consistency, and accuracy. PCR products give superior short to long (Table 1).
sequencing results when treated with ExoSAP-IT Express
reagent (Figure 1). Table 1. DNA recovery after purification.
Amplicon Agencourt™
ExoSAP-IT reagent
size AMPure™ XP beads
Superior quality scores 86 bp 10% 100%
103 bp 12% 100%
62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62 62
High signal intensity
545 bp 63% 100%
No background interference
1,007 bp 88% 100%
ExoSAP-IT Express reagent enables 100% recovery and provides effective cleanup of all amplicon
C A T G C A C C A T T C C T T G C G G C G G C G G T G C T C A A C G G C C T C A A C C sizes. In contrast, Agencourt AMPure XP beads were ineffective at purifying small amplicons, whether
No miscalls or N base calls determined by image analysis or by the Invitrogen™ Quant-iT ™ PicoGreen™ assay.

Figure 1. Sequencing results of a 1 kb PCR product treated with

ExoSAP-IT Express reagent. Treatment with ExoSAP-IT Express reagent
prior to sequencing eliminates miscalls and improves sequencing scores
(numbers and bars above sequence; quality score >60, probability of error
≤0.0001%). Sequence shown is approximately 400 bases from the primer
binding site.
Overview Also available in a high-throughput
ExoSAP-IT reagents are a proprietary mixture of formulation
exonuclease I combined with shrimp alkaline phosphatase
(SAP) in a specially formulated buffer that removes excess Applied Biosystems™ HT ExoSAP-IT™ Fast High-
primers and dNTPs following a PCR reaction (Figure 3). Throughput PCR Product Cleanup Reagent is an
Exonuclease I removes residual single-stranded primers alternative formulation of the original ExoSAP-IT reagent,
and any single-stranded DNA produced during PCR. SAP specifically designed for the unique requirements of
removes the remaining dNTPs from the PCR mixture that high-throughput, automated platforms and multichannel
may interfere with subsequent reactions. pipettes. HT ExoSAP-IT Fast reagent quickly provides high-
quality purified samples. This formulation offers decreased
Add ExoSAP-IT 37˚C, 4 min for treatment
Express reagent 80˚C, 1 min to inactivate viscosity and is ideal for automated liquid handling
platforms, with the same convenience and stability you
Intact PCR product have come to expect from ExoSAP-IT reagent.
Primers Exo I SAP
Double-
stranded
• 14 min cleanup protocol
DNA
+
Nucleosides • Stable at 4°C for 1 month and at room temperature for
Nucleotides +
Inorganic phosphate 2 days
(Pi)

• Scalable for PCR cleanup ranging from a single tube up

Figure 3. How ExoSAP-IT reagent works. Treat 5 μL of PCR product
with 2 μL of ExoSAP-IT reagent. Treatment is carried out at 37°C followed to a 384-well plate
by an incubation period at 80°C to completely inactivate both enzymes.
Once contaminants are removed, your PCR products are ready for
downstream applications such as Sanger sequencing, next-generation
sequencing (NGS), fragment analysis, single-nucleotide polymorphism
(SNP) analysis, in vitro transcription, or single-base extension.

The original ExoSAP-IT reagent formulation

Applied Biosystems™ ExoSAP-IT™ PCR Product Cleanup
Reagent has been cited in over 10,000 publications.
Thermo Fisher Scientific is the only manufacturer of
ExoSAP-IT reagents, assuring your lab of product
consistency and integrity.

• 30 min cleanup protocol

• Recommended for processing 1–96 samples at a time

• Best cost-per-reaction value

Which ExoSAP-IT PCR cleanup reagent is right for you?

ExoSAP-IT reagents offer a unique one-tube, one- with downstream applications. Achieve superior results
step enzymatic method for PCR cleanup. All ExoSAP- with ExoSAP-IT reagent—improve accuracy with higher
IT reagents provide 100% recovery of PCR products yields and full PCR product recovery.
regardless of the fragment sizes. This PCR cleanup method
removes excess primers and dNTPs and does not interfere Use our selection guide to determine which formulation is
best for your next experiment (Table 2).

Table 2. Selection guide for ExoSAP-IT reagents.

ExoSAP-IT reagent HT ExoSAP-IT Fast
ExoSAP-IT Express reagent
(original formulation) High-Throughput reagent
Protocol time 5 min 30 min 14 min
Format Single tube Single tube Single tube
8-tube strip 8-tube strip
96-well plate
Throughput level Low to high; recommended for Low to medium; High; recommended for
processing any sample size recommended for processing processing ≥96 samples at a time
1–96 samples at a time
Platform Single- or multichannel pipette, Single-channel pipette Automated liquid handling
automated liquid handling platforms (lower viscosity for
platforms better handling)
Freezes at –20°C No No Yes
Stability –20°C for up to 2 years –20°C for up to 2 years –20°C for up to 2 years; once
thawed, stable at 4°C for 1 month
and room temperature for 2 days
Ordering information
Product Quantity Cat. No.
100 reactions 75001.200.UL
480 reactions
75001.1.EA
(8-tube strips)
ExoSAP-IT Express PCR Product Cleanup Reagent 500 reactions 75001.1.ML
2,000 reactions 75001.4X.1.ML
5,000 reactions 75001.10.ML
100 reactions 78200.200.UL
500 reactions 78201.1.ML
ExoSAP-IT PCR Product Cleanup Reagent
2,000 reactions 78202.4X.1.ML
5,000 reactions 78205.10.ML
20 reactions 7859520RXN
1,000 reactions 785951000RXN
5,000 reactions 785955000RXN
HT ExoSAP-IT Fast High-Throughput 480 reactions
785951EA
PCR Product Cleanup Reagent (8-tube strips)
5,760 reactions
785951PK
(12 x 8-tube strips in a tray)
23,040 reactions
785954PK
(48 x 8-tube strips in a tray)

Find out more at thermofisher.com/exosapit

For Research Use Only. Not for use in diagnostic procedures. © 2020 Thermo Fisher Scientific Inc. All rights reserved. All trademarks
are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Agencourt and AMPure are trademarks of
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