The NGS DNA Fragmentation & Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality libraries for next generation sequencing. The kit uses intact genomic DNA (EDTA-free DNA or DNA resuspended in TE buffer) as input DNA without an additional DNA fragmentation step. Our technology provides a fast and simple workflow. DNA libraries can be generated around 2 hours with only 10 min hands-on time. Library multiplexing is possible.
NGS DNA Fragmentation & Library Prep Kit Workflow
The incorporation of DNA fragmentation in the kit makes it possible to directly use intact genomic DNA as input DNA without the need of mechanical DNA shearing or enzymatic DNA fragmentation. The NGS DNA Fragmentation & Library Prep Kit does not generate sequencing bias as compared to library using mechanical sheared DNA as input. Sequence coverage is also consistent between enzymatic shearing and mechanical shearing. The library size is inversely correlated with the incubation time of step 1 at 20°C.
Three index types are available for the kit of the illumina platform:
Non-index (Cat.# 30026): Libraries do not have index.
Index (Cat.# 30028): Each of the index primers contains a unique index sequence of 6 bases. Library multiplexing for 48 samples is possible. Index information can be downloaded here.
Unique dual index (Cat.# 30030): Library multiplexing for 96 samples is possible. With the unique features of our 4-Base Difference Index System, the index sequence is 8-base long and each index has 4 bases different from others. Our unique dual index primers effectively identify sequencing errors such as index hopping, mis-assignment of reads, and de-multiplexing errors etc. The unique dual index primers set consists of 96 pre-mixed unique pairs of index primers. Index information can be downloaded here.
Indexes are available for the MGI platform kits (Cat.# 34028).
Kit features:
Library conversion efficiency: 100 ng, 300 ng and 500 ng of intact genomic DNA were used as input.
The library size is inversely correlated with the incubation time of step 1 at 20°C.
NGS data comparison: enzymatic shearing versus mechanical shearing
Enzymatic shearing
• DNA shearing and library prep: BioDynami NGS DNA Fragmentation & Library Prep Kit
Mechanical shearing
• DNA shearing: Covaris sonication
• Library prep: BioDynami NGS DNA Library Prep Kit.
The NGS DNA Fragmentation & Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality libraries for next generation sequencing. The kit uses intact genomic DNA (EDTA-free DNA or DNA resuspended in TE buffer) as input DNA without an additional DNA fragmentation step. Our technology provides a fast and simple workflow. DNA libraries can be generated around 2 hours with only 10 min hands-on time. Library multiplexing is possible.
Usages:
For the growth of pathogenic and non-pathogenic microorganisms.
Principle:
Polypeptone provide carbon and nitrogen sources, vitamins and growth factors; Neisseria starch can promote the growth characteristics and enhanced hemolytic streptococci; sodium chloride to maintain osmotic balance; agar as culture medium coagulant.
Formulation (per liter):
Peptone: 10g
Beef extract powder :10g
Sodium chloride: 5g
Agar :15g
Final pH 7.4 ± 0.2
How to use:
1. Suspend 40g of the product, adding 1 L of distilled or deionized water, ,heated to boiling stirring until completely dissolved, dispensing into flask, 121 autoclaved 15min.
2.After the medium is cooled to about 50 , sterile procedures, basal medium was added per 100mL defibrinated sheep or rabbit blood 5-10mL, shake pour plate; also dispensing sterile tubes set into the slope. Spare.
3. take the bacteria to be tested fresh pure cultures inoculated crossed or coated on the plate, set the temperature of the incubator training requirements were cultured for a predetermined time.
4. Observe the results.
Quality control:
The following were inoculated after 36 ± 1 for 24h , the results are as follows:
bacteria name bacteria No. growth status colony characteristics
Escherichia coli ATCC25922 not good hemolysis
Staphylococcus aureus, ATCC6538 good β-hemolytic
Beta-hemolytic streptococcus CMCC (B) 32210 good β-hemolytic
Streptococcus pneumoniae CMCC (B) 31001 good α-hemolytic
Storage: Store in a dark, cool and dry place, tighten the cap immediately after use. Storage period of three years.
500g
As this is a 2 gene kit, we recommend purchase of 2 of the accompanying RT-qPCR master mix reagent: oasig Lyophilised OneStep RT-qPCR Master Mix 150 reactions.
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