BK Virus

The Bisulfite Sequencing Library Prep Kit (illumina platform) was developed for construction of NGS libraries using bisulfite treated DNA (50 ng – 500 ng) as input. DNA methylation is an epigenetic mechanism known to play a critical role in gene regulation and genomic imprinting by blocking transcription factor access to promoters and enhancers. Bisulfite sequencing is a popular technique in biomedical research based on C to T conversion under the treatment of sodium bisulfite.

Recently, NGS became a powerful tool to identify the DNA methylation status at the whole genome level with single-base resolution. However, it is well known that bisulfite treatment of the NGS libraries causes tremendous damage to the libraries.

Bisulfite-Seq kit comparison

In the case of the regular bisulfite seq library preparation (library prep before bisulfite conversion), the DNA shearing equipment and the expensive methylated adaptors are required. In addition, the subsequent bisulfite conversion causes tremendous DNA damage to the constructed libraries.

BioDynami has developed a unique library prep technology to solve the problems. The technology uses bisulfite treated DNA as input to avoid the significant library loss caused by bisulfite conversion. Furthermore, DNA shearing step and expensive methylated adaptors are not required with our kit. The DNA polymerase in the kit has high-fidelity amplification ability and uracil tolerance which is ideal for amplification of bisulfite sequencing libraries. The final library is strand specific.

Bisulfite Sequencing Library Prep Kit Workflow

Three index types are available for the kit:

Non-index (Cat.# 30091): Libraries do not have index.

Index (Cat.# 30092): Each primer contains a unique barcode sequence of 6 bases to identify the individual library. Library multiplexing capacity is up to 48 samples. Index information can be downloaded here.

Unique dual index (Cat.# 30093): The multiplexing of bisulfite sequencing library is up to 96 samples with unique dual indexes. We used a Four-Base Difference Index System to generate indexes that have at least 4 bases different from each other in the 8-base index. The index primers remove NGS errors including index cross-contamination, index hopping, reads mis-assignment etc. Index information can be downloaded here.

Kit advantages

• • • Easy and Quick protocol
      • Easy: Hands-on time only 10 minutes
      • Quick: Total protocol time around 1.5 hours
    • Simple workflow: Less steps
    • Directional library
    • Save magnetic beads more than 50%
    • Guaranteed high Bisulfite sequencing library conversion efficiency
    • Bisulfite treated DNA as input: From 50 ng to 500 ng

The Bisulfite Sequencing Library Prep Kit (illumina platform) was developed for construction of NGS libraries using bisulfite treated DNA (50 ng – 500 ng) as input. DNA methylation is an epigenetic mechanism known to play a critical role in gene regulation and genomic imprinting by blocking transcription factor access to promoters and enhancers. Bisulfite sequencing is a popular technique in biomedical research based on C to T conversion under the treatment of sodium bisulfite.

Overview

• Sequentially purify total RNA (and miRNA), DNA and proteins from a single sample
• No sample splitting or need to use phenol or precipitation methods
• Purify RNA/DNA/Protein from cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

RNA/DNA/Protein Purification Plus Kit

This kit provides a rapid method for the high throughput isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, or yeast. The kit employs two columns: 1) for gDNA purification and 2) for RNA purification utilizing Norgen’s resin columns (superior for the binding of all RNA sizes including miRNA). The proteins are also purified on the second column after RNA elution. The proteins are eluted in buffer and are ready for downstream application without any further clean up required. The proteins can be quantified directly, used in western blots, ELISA or mass spectrometry. This kit provides a rapid spin-column method for the isolation and purification of total RNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants.

RNA/DNA/Protein Purification Plus Micro Kit

This kit provides a rapid spin-column method for the isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, microdissected samples including LCM, stem cells, sorted cells, and CTC. The total RNA, genomic DNA and proteins are all column purified in less than 30 minutes. The RNA and DNA can be eluted in as little as 20 µL while the protein can be eluted in as little as 50 µL. This kit provides the same performance as if the samples were isolated from dedicated kits.

RNA/DNA/Protein Purification 96-Well Plus Kit

The kit employs two plates: 1) for DNA purification and 2) for RNA purification utilizing Norgen’s resin (superior for the binding of all RNA sizes including miRNA). Please see the protocol schematic below.

Details

Supporting Data

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Kit Specifications
Binding Capacity Per Well	50 μg for RNA 20 μg for DNA 150 μg for protein
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Size of DNA Purified	≥ 30 kb
Time to Complete 10 Purifications	35 minutes
Maximum Amount of Starting Material:Animal CellsAnimal TissuesBloodBacteriaYeastFungiPlant Tissues	1 x 106 cells10 mg100 µL1 x 109 cells1 x 108 cells40 mg40 mg
Average Yield* Liver (5 mg)	12.5 μg RNA 2 μg DNa  55 μg Protein

* average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.

Storage Conditions and Product Stability
The Protein Loading Dye should be stored at -20°C after the addition of DL-Dithiothreitol (DTT).

Component	Cat. 47700 (50 preps)	Cat. 51600 (50 preps)	Cat. 51700 (96 preps)
Buffer SKP	40 mL	40 mL	–
Lysis Buffer Q	–	–	40 mL
Wash Solution A	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL
Elution Solution A	6 mL	6 mL	20 mL
Elution Buffer F	15 mL	15 mL	2 x 15 mL
Wash Solution C	30 mL	30 mL	60 mL
Binding Buffer A	8 mL	8 mL	8 mL
Elution Buffer C	8 mL	8 mL	30 mL
Protein Neutralizer	4 mL	4 mL	4 mL
Protein Loading Dye	2 mL	2 mL	3 x 2 mL
gDNA Purification Columns	50	–	–
gDNA Purification Micro Columns	–	50	–
gDNA Purification 96-Well Plate	–	–	1
RNA/Protein Purification Columns	50	–	–
RNA/Protein Purification Micro Columns	–	50	–
RNA/Protein Purification 96-Well Plate	–	–	1
Collection Tubes	150	150	–
Collection Plate	–	–	5
Elution Tubes (1.7 mL)	150	150	–
Elution Plate	–	–	3
Lysis Preparation Plate	–	–	2
Adhesive Tape	–	–	4
Product Insert	1	1	1

Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Primer and probe mix (150 reactions)
Reverse Transcription, target specific primers (RNA genome viruses only)
Copy number standard curve (sufficient for multiple standard curves)
Internal extraction control – Read through VIC channel*
Endogenous control (150 tests)
RNAse/DNAse free water
*alternative fluorophores available on request