BKV/JCV TaqMan PCR Detection Kits

Description

Attogene has developed the first of its kind and patent pending technology that uses a novel RNA/DNA substrate tagged on the 5′ and 3′ end with a Biotin that is attached to our streptavidin colloidal gold reporter molecules and a test line tag. In the absence of RNases H, the gold tagged RNA/DNA molecule will flow up the lateral flow strip and bind to the test line using an anti tag antibody (INDICATING that NO RNase H is detected). When RNases H is present or added, however, the RNA strand of the RNA/DNA substrate is cleaved, and the Biotin tagged 5′ and 3′ tags are spatially separated in solution cauing the test line to disapear.

  This test can be used to rapidly and efficiently detect ribonucleases H activity and is a perfect tool for monitoring unit activity or residual RNAse H activity.  

RNase H activity in a convenient and sensitive colormetric assay that delivers results in real time. Great for Quality Testing for RNase H activity in enzyme preparations and determination of RNase H unit activity.

• HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) or allele-specific amplification (ASA).Please note: This DNA polymerase is also available as a nuclease deficient variant, featuring higher robustness towards potential PCR inhibitors and compatibility with real-time dyes such as our GreenDye.Benchmarking with products of competitors conducted by us and others show that the HiDi® DNA polymerase family is the first choice for highly selective PCRs, such as genotyping by allele-specific PCR, HLA genotyping, analysis of single CpG methylation sites or the detection of mutations in a high background of wild-type sequences. By using HiDi® Taq DNA polymerase, less than 10 copies of a mutation can be detected in a background of >10.000 wild-type copies straight away without any other tedious assay optimization.HiDi® Taq DNA polymerase harbours a nuclease function and therefor is also suitable for use with hydrolysis probes (TaqMan® probes etc.). It has also been shown that HiDi® DNA polymerase family is highly suitable for quality control and mutation identification in CRISPR-Cas or TALEN-based applications.Several independently conducted studies show that HiDi® Taq DNA polymerase is ideally suited for use in asPCR in numerous research areas ranging from mutation detection to genome editing. (read more)
  For research use and further manufacturing.In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.

HiDi is available as:

HiDi® DNA Polymerase (>>)
HiDi® Taq DNA Polymerase (>>)
HiDi® 2x PCR Master Mix (>>)
HiDi® Taq 2x PCR Master Mix (>>)

Casestudies:
HiDi® DNA Polymerase: Applications from mutation detection to genome editing (read more)

Example Primer Design

Matching vs. mismatching nucleotide is placed at the 3′-end of the primer for best discrimination results.

Example Results – There´s no accounting for taste

Cilantro: some people love it in their food, some hate it. Here we are detecting a genomic SNP (rs72921001) in HeLa genomic DNA. This SNP is reported to be close to a number of genes coding for olfactory receptors. (Reference: Eriksson N. et al. (2012), “A genetic variant near olfactory receptor genes influences cilantro preference.”)

Considering, that only the C-allele specific primer is extended and yielding in a specific amplicon, we can conclude a genetic predisposition in disliking cilantro, as this SNP is significantly associated with detecting a soapy taste to cilantro.  

Allele-specific PCRs were performed from 1 ng/µl of HeLa gDNA in the presence of a realtime dye, indicating the amplification of the C-allele specific primer only. The A-allele specific primer is discriminated, thus not amplified up to 50 cycles.

PCR products were subsequently analysed on a 2.5% agarose gel. Specific product is visualized by ethidium bromide staining at the amplicon length of 109 bp.

HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) or allele-specific amplification (ASA).

Please note: This DNA polymerase is also available as a nuclease deficient variant, featuring higher robustness towards potential PCR inhibitors and compatibility with real-time dyes such as our GreenDye.

Ideal magnet for all of your low volume needs. From Low elution 96-well PCR applications to 384-well, the X396 has  you covered. No need to purchase two separate plates anymore. 96-well volumes as low as 5 µL are achieved from 96-well, PCR plates on the inside of the magnet, while the outside of each magnet will pull beads to well sides in 384-well plates.

ANSI/ SBS Footprint (127.75mm x 85.50mm) to fit into any automated liquid handling robot on bottom. ANSI/SBS footprint on top to accept all common microplates.

Integrated Cushion base for maximum recovery. Helps aid in set-up, robot positioning inconsistencies, and labware consumable inconsistencies

Features include solid aluminum alloy construction and hard coat anodized finish for years of trouble-free use, and compatible with any magnetic beads

SKU #

A040523

Features

• ANSI/ SBS footprint
• Compatible with 384-well and 96-well plates
• Solid Aluminum alloy design
• Precision manufactured for more consistent results
• Fast magnetic bead separations
• Made in USA 
• For Research use only

Compatibility

• 384-well pcr plates
• Some 384-well deep well plates
• Full-, Semi-, and un-Skirted 96-well PCR Plates
• Any magnetic beads
• Any common liquid handlers i.e. Tecan®, Opentrons®, Hamilton®, Agilent®, Beckman®, etc.

Volumes

384-Well PCR

Maximum – 39 µL 

Minimum – 5 µL 

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96-Well PCR

Maximum – 60 µL (0.2 mL for some beads)

Minimum – 4 µL

Ideal magnet for all of your low volume needs. From Low elution 96-well PCR applications to 384-well, the X396 has you covered. No need to purchase two separate plates anymore. 96-well volumes as low as 5 µL are achieved from 96-well, PCR plates on the inside of the magnet, while the outside of each magnet will pull beads to well sides in 384-well plates.