PCRbeam™ Fast PCR Detection Kit

Opis towaru

highQu PCRbeam™ Fast PCR Detection Kit is a convenient tool for fast detection

of gene-specific amplification products obtained by PCR, LAMP or RPA.

The detection is based on immunological reaction driven by Biotin and FITC (fluorescein isothiocyanate),

thus the amplified DNA shall include Biotin and FITC labels. The PCR amplification has to be performed

with one primer labeled with FITC at 5’-end and one primer labeled with Biotin at 5’-end.

Alternatively the use of one of the labeled primers can be replaced by gene-specific FITC

or Biotin labeled probe. Kit includes PCRbeam™ Membrane Strips that are coated with biotin-ligand

on the test band and an anti-rabbit antibody on the control band. The bottom part of the strip which

is used for sample application contains an anti-FITC antibody attached to gold particles.

PCRbeam™ Detection Buffer is Tris-buffered saline enabling the detection.

The PCRbeam™ Fast PCR Detection Kit can be applied for established tests or home-brew assays

as a fast and sensitive yes/no detection method. The detection sensitivity is up to 100 fold higher

than the one achievable with ethidium bromide stained gels what provides an environment friendly save

and economical alternative to the use of mutagen stains.

For establishing sensitive PCR-based tests before PCRbeam™ detection we recommend the use

of hot-start PCR enzymes or master mixes, like highQu ALLin™ Hot Start Taq Mastermix or ALLin™ Hot Start Taq DNA Polymerase.

Principle: The membrane strip is soaked for 10 minutes into the vial with the detection buffer mixed with PCR product.

The lateral sample flow driven by gold particles moves the solution up the strip. FITC labeled DNA strand binds

with the anti-FITC antibody on the gold particle and Biotin labeled DNA strand is caught by Biotin ligand attached to the test band.

As both DNA strands remain hybridized at room temperature, the test band builds an aggregate that develops red-blue color.

Excess gold particles that were not caught by FITC move up the strip and the anti-FITC antibody binds to the anti-rabbit antibody

to develop the red-blue colored control band.

If there is no PCR product in the reaction, then only the control band will be visible.

If there is a specific product, the test band will be colored as well.

Applications

Low throughput PCR, LAMP, RPA based tests.

Sensitive detection of specific amplification products.

Fast and 20x more sensitive alternative to EtBr stained gels.

Economical alternative to qPCR-based detection.

Benefits

Sensitive detection of PCR, LAMP, RPA gene-specific products.

No gel loading after PCR, no ethidium bromide handling.

Saved costs compared to qPCR-based detection methods.

Fast and easy procedure with little hands on time.