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Multiomic ATAC-Seq Amplification Kit

BD Rhapsody™ Multiomic ATAC-Seq Amplification Kit

(RUO)
Product Details
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BD Rhapsody™
Human,Mouse (Reported)
Single cell multiomics experiments on the BD Rhapsody™ platform (Tested During Development)
RUO


Description

This kit provides the necessary reagents to support the preparation of single-cell multiomic ATAC-Seq + Whole Transcriptome Analysis libraries from the nuclei prepared using the BD Rhapsody™ ATAC-Seq Tagmentation and Supplemental Reagents Kit (sold separately) and captured on the BD Rhapsody™ Single-Cell Analysis system. For detailed instructions on the tagmentation step and single nuclei capture on the BD Rhapsody™ Single-Cell Analysis system or the BD Rhapsody™ HT Xpress system, refer to the protocols listed in the Recommended Assay Procedure section. Optimized for use with Rhapsody Enhanced Cell Capture Beads V3, the BD Rhapsody™ Multiomic ATAC-Seq Amplification Kit provides all the reagents required for the ligation step performed on the transposed fragments captured from the nuclei loaded onto the BD Rhapsody™ Single-Cell Analysis system. This kit also provides a set of ready-to-use index primers to prepare 8 unique single-indexed libraries from the DNA fragments denatured off the beads after the reverse transcription and Exonuclease I treatment. For detailed instructions on the ligation, reverse transcription, Exonuclease I treatment, and library preparation steps, refer to the protocol listed in the Recommended Assay Procedure section.

Recommended Assay Procedures

After retrieving and washing the Cell Capture Beads from the Rhapsody Cartridge, a ligation mix, prepared using the reagents provided in the kit, is added to the Beads to ligate the double-stranded DNA fragments to the bead oligos. Following the ligation step, a reverse transcription mix, consisting of the reverse transcriptase and all the required buffers and dNTPs, is prepared and added to the beads to perform the reverse transcription of the mRNA fragments captured on the dT oligos on the bead. At the same time, the reverse transcription serves as a gap-filling treatment to repair the tagmentase-induced nicks in the transposed DNA fragments ligated to the beads.

For detailed instructions on the tagmentation step, splint-bead generation, ligation, reverse transcription, Exonuclease I treatment, and preparation of multiomic ATAC-Seq + Whole Transcriptome Analysis libraries, refer to Single-Cell ATAC and mRNA Whole Transcriptome Analysis (WTA) Library Preparation Protocol (Doc ID 23-24474).

For instructions on how to use the BD Rhapsody™ Single-Cell Analysis system, refer to the BD Rhapsody™ HT Xpress Single-Cell Analysis System Instrument User Guide (Doc ID 23-24257) or the BD Rhapsody™ HT Xpress System Instrument User Guide for Scanner-free workflow (Doc ID 23-24256).

For sample preparation prior to the single cell ATAC-Seq assay, refer to BD Rhapsody™ System Preparing Single Cell Suspensions Protocol (Doc ID 23-24126).

For information on the bioinformatic pipeline for the ATAC-Seq assay, refer to BD® Single-Cell Multiomics Bioinformatics Handbook (Doc ID 54169).

For safety information, see the BD Rhapsody ™ Single-Cell Analysis System Instrument User Guide (Doc ID 23-24257).

Use standard laboratory safety protocols. Read and understand the safety data sheets (SDSs) before handling chemicals. To obtain SDSs, go to regdocs.bd.com or contact BD Biosciences technical support at scomix@bdscomix.bd.com.

Warning: All biological specimens and materials coming into contact with them are considered biohazardous. Handle as if capable of transmitting infection and dispose of with proper precautions in accordance with federal, state, and local regulations. Never pipette by mouth. Wear suitable protective clothing, eyewear, and gloves.

571361 Rhapsody ATAC MO Amplification Kit, Kit component 51-9024048 PCR MasterMix/Watch Maker Buffer

Danger: PCR Master Mix contains 5-10 % Tris base, CAS number 77-86-1, EC number 201-064-4 and 1-5 % tetramethylammonium chloride, CAS number 75-57-0, EC number 200-880-8. These reagents are classified as hazardous according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS).

Danger

Causes skin irritation.

Cause damage to organs (Central Nervous System) via ingestion.

Harmful if swallowed.

Prevention

Wash skin thoroughly after handling

Wear protective gloves/protective clothing/eye protection/face protection/hearing protection

Do not breathe dust/fume/gas/mist/vapors/spray

Do not eat, drink or smoke when using this product

Response

IF ON SKIN: Wash with plenty of soap and water.

Take off contaminated clothing and wash it before reuse.

If skin irritation occurs: Get medical advice/attention.

Specific treatment (see Safety Data Sheet)

IF SWALLOWED: Call a POISON CENTER or doctor/physician if you feel unwell

Rinse mouth

IF exposed or concerned: Call a POISON CENTER or physician

Disposal

Store locked up.

Dispose of contents/container to an approved facility in accordance with local, regional, national and international regulations.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  3. For U.S. patents that may apply, see bd.com/patents.
  4. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
571361 Rev. 1
Components
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Description Quantity/Size Part Number EntrezGene ID
Ligation Buffer N/A 51-9023094 N/A
Ligase N/A 51-9023093 N/A
Nuclease Free Water N/A 51-9023086 N/A
ATAC-Seq Library Forward Primer N/A 51-9023097 N/A
ATAC-Seq Library Reverse Primer 1 N/A 51-9023098 N/A
ATAC-Seq Library Reverse Primer 2 N/A 51-9023099 N/A
ATAC-Seq Library Reverse Primer 3 N/A 51-9023100 N/A
ATAC-Seq Library Reverse Primer 4 N/A 51-9023101 N/A
ATAC-Seq Library Reverse Primer 5 N/A 51-9023102 N/A
ATAC-Seq Library Reverse Primer 6 N/A 51-9023103 N/A
ATAC-Seq Library Reverse Primer 7 N/A 51-9023108 N/A
ATAC-Seq Library Reverse Primer 8 N/A 51-9023109 N/A
RNase Inhibitor N/A 51-9024039 N/A
0.1M DTT, Molecular Biology Grade 1 Each (1 ea) 51-9022688 N/A
Bead Resuspension Buffer N/A 51-9023126 N/A
Elution Buffer N/A 51-9023107 N/A
Splint oligo Removal Buffer N/A 51-9024041 N/A
PCR MasterMix N/A 51-9024048 N/A
571361 Rev. 1

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.