STRboost™ Protocol for enhanced PCR with difficult samples

Product Nos. 63401-011, 63401-001, 63400-001

Product Description

STRboost is a novel reagent that substantially enhances multiplex PCR amplification reactions used for human identification applications, resulting in improved accuracy, sensitivity and precision. Including STRboost in amplification reactions helps resolve stochastic effects typically associated with sample degradation or inhibition (e.g. peak balance issues, allele dropout, and suboptimal peak height definition). STRboost is ideal for generating DNA profiles from difficult samples, such as those containing inhibitors (e.g. humic acid, hemetin), degraded samples, or derived from limited sources (e.g. trace evidence and low copy number).

STRboost is easy to incorporate into existing workflows. Analysts can continue use of their own cycling conditions and equipment. The reagent is compatible with commercially available genetic identification kits such as ABI’s Identifiler™ and Promega’s PowerPlex 16™ Systems and does not interfere with automated detection systems (e.g. ABI 3100 PRISM Genetic Analyzer).

Precautions and Disclaimer

This product is for R&D use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices. 

Storage/Stability

Store at room temperature. Use within 6 months of purchase date for optimal product performance.

Procedure

When using STRboost, there is no change to your standard multiplex STR analysis protocol. Simply add STRboost to amplification reactions.

The following is an example of a multiplex PCR amplification reaction containing STRboost using the AmpFℓSTR® Identifiler kit (ABI). The resulting PCR product can be used directly in downstream applications without further purification.

Example: Multiplex PCR Amplification Reaction
Genomic DNA (1 ng) 5 μl
PCR reaction mix 10 μl
Primer F set 5 μl
AmpliTaq® Gold polymerase 0.5 μl
STRboost 2.5 μl
Water + 2.5 μl
FINAL VOLUME 25 μl

Example: PCR Cycling Protocol
95˚C 11 min
94˚C 1 min
59˚C 1 min
72˚C 1 min
60˚C 60 min
4˚C ∞

Example: Sample preparation for ABI PRISM 310 Genetic Analyzer
1. Combine the necessary amount of the following regents in a single microfuge tube as shown:

(number of samples+2) x 24.5 μl Hi-Di Formamide
(number of samples+2) x 0.5 μl GeneScan-500 LIZ Size Standard

2. Vortex the tube to mix. Spin the tube briefly in a microcentrifuge.
3. Aliquot 25 μl of Hi-Di Formamide/GeneScan-500 LIZ solution into Genetic Analyzer sample tubes.
4. Add 1.5 μl PCR product containing STRboost per tube.
5. Mix by pipetting up and down.
6. Seal each tube with a septum.
7. Continue with standard sample preparation protocol.

Samples can be used with all the major human identification kits, including:
AmpFℓSTR® Identifiler PCR Amplification Kit1
AmpFℓSTR® Profiler Plus® ID PCR Amplification Kit1
AmpFℓSTR® Profiler Plus® COfiler® PCR Amplification Kit1
AmpFℓSTR® MiniFiler™ PCR Amplification Kit1 AmpFℓSTR® YFiler® PCR Amplification Kit1
PowerPlex® 16 System2
PowerPlex® Y System2
GenePrint® Fluorescent STR Systems FFFL Multiplex2

Materials

     
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