The NGS Low Input DNA Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality NGS libraries with low input DNA amount from 1 ng to 400 ng. The kit allows scientist to study samples with limited DNA such as tumor samples, patient samples, and other specially collected samples (FACS sorting etc.). The kit has high library conversion efficiency with as little as 1 ng DNA input. The fast and simple 1.5-hour protocol makes libraries with even coverage and low GC-bias based our unique chemistry for DNA end-polishing and ligation.
The kit needs double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library preparation, and is compatible with DNA fragments generated from both enzymatic approach (for example, BioDynami DNA fragmentation enzymes, Cat.# 40061 and Cat.# 40062) and mechanical approaches such as sonication and nebulization etc.
NGS Low Input DNA Library Prep Kit Workflow
Three index types are available for the NGS Low Input DNA Library Prep Kit of illumina platform:
Non-index (Cat.# 30022): Libraries do not have index.
Index (Cat.# 30024): Each of our index primers contains a unique barcode sequence with 6 bases that can be used to identify the low input DNA libraries. Library multiplexing up to 48 samples is possible. Index information can be downloaded here.
Unique dual index (Cat.# 30025): Library multiplexing up to 96 low input DNA libraries is possible with unique dual indexes with our Four-Base Difference Index System. The system allows us to make indexes for the libraries that have at least 4 bases different from each other in the 8-base barcode length. Our unique dual index primers can reduce sequencing errors such as de-multiplexing errors, amplification errors, mis-assignment of reads, index cross-contamination, and also index hopping. The kit includes 96 pre-mixed unique pairs of index primers. Index information can be downloaded here.
Indexes are available for the MGI platform kits (Cat.# 34024).
Kit advantages:
Comparison of library conversion efficiency under the same NGS library preparation condition. Input DNA amounts are 1 ng and 10 ng, respectively. DNA was mechanical sheared with Covaris before library prep. BioDynami kit: NGS Low Input DNA Library Prep Kit (Cat. #30022).
Comparison of library yield under the same NGS library prep condition. Input DNA amounts are 1 ng, 10 ng, and 100 ng. DNA was mechanical sheared with Covaris before library prep. BioDynami kit (Cat. #30022). PCR cycle numbers were indicated.
The NGS Low Input DNA Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality NGS libraries with low input DNA amount from 1 ng to 400 ng. The kit allows scientist to study samples with limited DNA such as tumor samples, patient samples, and other specially collected samples (FACS sorting etc.). The kit has high library conversion efficiency with as little as 1 ng DNA input. The fast and simple 1.5-hour protocol makes libraries with even coverage and low GC-bias based our unique chemistry for DNA end-polishing and ligation.
This kit provides a rapid method for the isolation and purification of both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. The kit can be used to isolate all sizes of RNA from the cytoplasmic and nuclear RNA fractions, including all small RNA species without any requirement for phenol. Included in the kit are sufficient reagents to perform either 50 cytoplasmic RNA preparations or 25 cytoplasmic and 25 nuclear RNA preparations. Ten samples can be processed in approximately 45 minutes. This kit is also available in a 100 prep size.
Background
In certain circumstances it is desirable to be able to isolate fractionated RNA as opposed to total RNA. For example, it may be preferable to isolate only mature, cytoplasmic RNA for some studies on expression profiling. Alternatively it may be desirable to isolate nuclear RNA in order to investigate and study pre-processed (non-spliced) RNA. Furthermore, this kit can be used to isolate RNA for downstream applications where it is necessary to avoid DNA contamination, since the cytoplasmic fraction has been shown to be free of all traces of genomic DNA.
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| Kit Specifications | |
| Maximum Column Binding Capacity | Up to 50 µg RNA |
| Maximum Column Loading Volume | 650 µL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Time to Complete 10 Purifications | 45 minutes |
| RNA Yield HeLa (1 x 106) – Cytoplasmic RNA HeLa (1 x 106) – Nuclear RNA | 15 µg ≤ 3.5 µg |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
| Component | Cat. 21000 (50 preps) | Cat. 37400 (100 preps) |
|---|---|---|
| Lysis Buffer J | 20 mL | 2 x 20 mL |
| Buffer SK | 40 mL | 2 x 40 mL |
| Wash Solution A | 38 mL | 2 x 38 mL |
| Elution Buffer E | 6 mL | 2 x 6 mL |
| Mini Spin Columns | 50 | 100 |
| Collection Tubes | 50 | 100 |
| Elution Tubes (1.7 mL) | 50 | 100 |
| Product Insert | 1 | 1 |
Opentrons Flex 1-Channel pipettes enable highly accurate pipetting, optimized for automation.
Flex pipettes use air displacement technology to offer highly accurate pipetting with pipette volume ranges from 1 to 50 µL and 5 to 1000 μL. Smart sensors support automatic calibration, real-time positioning, and error detection. Choose the pipettes that fit your workflow — the gantry supports any two Flex 1-Channel or 8-Channel Pipettes — and swap them out yourself when your workflow changes.
Opentrons Flex 1-Channel pipettes enable highly accurate pipetting, optimized for automation.
Flex pipettes use air displacement technology to offer highly accurate pipetting with pipette volume ranges from 1 to 50 µL and 5 to 1000 μL. Smart sensors support automatic calibration, real-time positioning, and error detection. Choose the pipettes that fit your workflow — the gantry supports any two Flex 1-Channel or 8-Channel Pipettes — and swap them out yourself when your workflow changes.