TDL5E Bench top low speed refrigerated centrifuge machine 

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The Heat&Run® gDNA removal kit removes contaminating gDNA from RNA prior to RT-qPCR.

The kit is based on the recombinant heat labile dsDNase, which is irreversibly inactivated at low temperatures (5 minutes at 58ºC or 15 minutes at 55ºC).

Reverse Transcription can be performed in the same tube, thereby minimizing pipetting steps and reducing hands-on time.

Protocol

The kit contains HL-dsDNase which efficiently removes gDNA from RNA preps before running RT-qPCR.

• gDNA is removed, leaving RNA ready for reverse transcription in the same tube (Figure 1).
• Heat-labile dsDNase can easily be inactivated.
• This procedure minimizes pipetting steps and reduces hands-on time.
• The Heat&Run kit is especially well suited for high throughput experiments.

The high activity of the HL-dsDNase at low temperature and its heat-lability makes it ideal to use in a combination with a heat activated RT enzyme. The contaminating genomic DNA is cleaved at ambient temperature and increasing the temperature over 50°C will inactivate the DNase and start the Reverse Transcriptase.

Do you require gDNA removal in applications other than RT-qPCR? Contact our support team for assistance in implementing dsDNase treatment in your workflow.

Kit Contents

• 10X reaction Buffer
• HL-dsDNase (50 or 250 reactions)

The NGS FFPE DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality FFPE DNA libraries using 10 ng to 400 ng of input DNA isolated from formalin-fixed, paraffin-embedded (FFPE) samples. The DNA damage caused by fixation makes it difficult to construct high quality libraries. Our kit has been optimized to repair damaged DNA in the reactions. Multiplexing of the NGS FFPE DNA Library is possible.

NGS FFPE DNA Library Prep Kit Workflow

FFPE samples are a great resource for biomedical research. However, the methods for fixation and condition of storage significantly damage the DNA in the samples.  Thus, the extraction of high quality DNA from FFPE samples is often a challenge.  Low yield and low quality of FFPE DNA usually are common because of the limited tissue material and the DNA degradation.

As a result, it is usually difficult to construct high quality NGS libraries from low amount and low quality of FFPE DNA. In order to address this issue, we have developed the NGS FFPE DNA Library Prep Kit to make high quality libraries from the low input of FFPE DNA samples.

Three index types are available for the NGS FFPE DNA Library Prep Kit of the illumina platform:

Non-index (Cat.# 30035): Libraries do not have index.

Index (Cat.# 30037): Each of our index primers contains a unique barcode DNA (6 bases long) that can be used to identify individual library. Multiplexing of libraries is up to 48 samples. Index information can be downloaded here.

Unique dual index (Cat.# 30039): FFPE DNA library multiplexing is possible with 96 samples based on the unique dual indexing system. Our unique Four–Base Difference Index System allows us to make indexes that have at least 4 bases different from each other in the 8-base index sequence. Our unique dual indexing primers can effectively remove NGS errors including index hopping, de-multiplexing errors, index cross-contamination, mis-assignments etc. The unique dual index primer set includes 96 pre-mixed unique pairs of i5 and i7 index primers. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34037).

Kit advantages:

• Fast and simple protocol
  • Making libraries in just 1.5 hours
  • 10 minutes of hands-on time
• Easy procedure
  • Ready-to-use master mix to reduce the tedious mixing
  • Less reaction components to simplify the reaction setup
• Less magnetic beads needed for the purification steps: saving more than 50% of the expensive beads
• Low input FFPE DNA: From 10 ng to 400 ng

Comparison of library conversion efficiency under the same condition. Input DNA amount is 25 ng.

Comparison of library yield under the same condition. Input DNA amount is 25 ng.

About

A universal PCR master mix for allele-specific PCR assays. Precision fluorescent signal generation with consistently high performance at any reaction volume.

PACE Genotyping Master Mix uses a novel, universal, fluorescent reporting cassette to produce machine-readable fluorescent signals corresponding to genotypes. PACE compatible genotyping assays are comprised of two competitive allele specific forward primers (which differ in their terminal 3’ bases and unique 5’ tail sequences) and a common, reverse primer.  PACE Genotyping Master Mix is supplied at 2x concentration and with ROX normalising dye at a range of levels to ensure compatibility with your qPCR instrument or reader.

Genotyping assay designs are available from 3CR Bioscience through our free PACE assay-design service; once designed, users can purchase assay primers independently or through 3CR Bioscience using our partial or full-assay validation service. PACE Genotyping Master Mix is also compatible with KASP™ and Amplifluor® marker assays.

REQUIRED COMPONENTS

• qPCR machine or Thermocycler + Fluorescent plate reader
• PCR plate or equivalent and appropriate optically clear seal
• Template DNA
• PCR-grade water
• Genotyping assays