Each sample that will be multiplexed will require the MULTIseq anchor oligos and a unique MULTIseq barcode. The common Anchor/Co-anchor Lipid Modified Oligos (LMOs) are components of the MULTIseq LMO001 kit. Each sample will also need a unique MULTIseq barcode oligo that can be custom ordered by sending an email to dnaoligo@milliporesigma.com. There are two types of MULTIseq barcode oligos, one for 3’ capture and one for 5’ capture. Each barcode oligo contains an 8bp variable region. The 8bp sequence will be provided when ordered. The MULTIseq Additive Primer, NGS (Next Gen Sequencing) prep oligos and all oligos that will be needed to complete the scRNAseq process can also be custom ordered by email or through SigmaAldrich.com/nextgenoligos.
Przejdź do
LMO001
MULTI-seq Oligo modyfikowane lipidami
for Single Cell and Single Nucleus Multiplexing
Synonim(y):
LMO, Oligole modyfikowane lipidami
Wybierz wielkość
| Rozmiar/SKU | Dostępność | Cena netto |
|---|---|---|
30 reactions | Skontaktuj się z Obsługą Klienta, aby uzyskać informacje na temat dostępności | 2150,00 zł |
100 reactions | Skontaktuj się z Obsługą Klienta, aby uzyskać informacje na temat dostępności | 5120,00 zł |
Informacje o tej pozycji
2150,00 zł
Quality Segment
form
liquid
usage
sufficient for 100 sample(s) (100RXN kit is adequate for labelling 100 samples when using 1 μL of each LMO per sample), sufficient for 30 sample(s) (30RXN kit is adequate for labelling 30 samples when using 1 μL of each LMO per sample)
concentration
50 μM (each)
technique(s)
PCR: suitable, multiplexing: suitable (Single Cell and Single Nucleus)
shipped in
dry ice
storage temp.
−20°C
General description
Features and Benefits
- Efektywność. MULTI-seq zwiększa przepustowość i zmniejsza koszty odczynników sekwencjonowania RNA opartego na kropelkach.
- Elastyczność. Kompatybilny z dowolną platformą generowania kropli, system indeksowania umożliwia użytkownikom jednoczesne uruchamianie i analizowanie do 96 próbek z kodem kreskowym w 8-pasmowym urządzeniu kropelkowym.
- Wyraźniejsze wyniki. Indeksy zapewniają wyższą jakość danych w porównaniu z nieindeksowanymi metodami analizy w postaci identyfikacji dubletów i zatrzymywania danych z komórek o niskiej zawartości RNA.
Other Notes
- Ten produkt jest przeznaczony wyłącznie do użytku badawczo-rozwojowego. Nie jest przeznaczony do leków, użytku domowego ani innych zastosowań.
- Unikalne kody kreskowe oligo nie są dołączone i należy je zakupić osobno.
- Kod kreskowy 3' oligo z ogonem poly-A do wzbogacania mRNA: 5′-CCTTGGCACCCGAGAATTCCA-8-base index-A30-3′
- Kod kreskowy oligo do syntezy 5' biblioteki cDNA: 5'-CCTTGGCACCCGAGAATTCCA-8-base indexCCCATATAAGAA-3'
- Ponadto, 96 x 5′-końcowych i 96 x 3′-końcowych unikalnych kodów kreskowych oligo jest dostępnych do nabycia oddzielnie. Szczegóły dotyczące naszego niestandardowego produktu kodów kreskowych, Next-Gen Sequencing Oligos (NGSO), można znaleźć na stronie SigmaAldrich.com/nextgenoligos. Aby zminimalizować zanieczyszczenie krzyżowe starterów kodów kreskowych, zamów NGSO-Silver lub najlepiej NGSO-Gold. Prosimy o zapoznanie się z dostępnymi specyfikacjami, a następnie przesłanie zapytania ofertowego dotyczącego kodów kreskowych MULTI-seq na adres dnaoligos@milliporesigma.com. Sekwencje nie są dostarczane przed zakupem, ale zostaną uwzględnione w dokumentacji produktu w momencie dostawy.
Reagents Provided
- LMO001A Lignoceric Anchor with DNA Oligo
- LMO001B Palmitic Co-anchor with DNA Oligo
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Ta pozycja | |||
|---|---|---|---|
| description for Single Cell and Single Nucleus Multiplexing | description Design and Order Custom Oligos | description - | description - |
| Quality Level 200 | Quality Level - | Quality Level 200 | Quality Level 100 |
| form liquid | form - | form - | form - |
| shipped in dry ice | shipped in - | shipped in wet ice | shipped in - |
| storage temp. −20°C | storage temp. - | storage temp. 20-25°C | storage temp. - |
| concentration 50 μM (each) | concentration - | concentration - | concentration - |
| technique(s) PCR: suitable, multiplexing: suitable (Single Cell and Single Nucleus) | technique(s) - | technique(s) whole genome amplification: suitable | technique(s) - |
Klasa składowania
12 - Non Combustible Liquids
Temperatura zapłonu (°F)
Not applicable
Temperatura zapłonu (°C)
Not applicable
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What is needed to add MULTIseq to my scRNAseq workflow?
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How do I order the 8-bp index sample barcodes?
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The 8 bp index is a component of the MULTI-seq barcode. There are two types of MULTIseq barcode oligos, one for 3’ capture and one for 5’ capture. Each barcode oligo contains an 8 bp variable region. The 8 bp sequence will be provided when ordered. The MULTIseq Additive Primer, NGS (Next Gen Sequencing) prep oligos and all oligos that will be needed to complete the scRNAseq process can also be custom ordered by email (dnaoligo@milliporesigma.com) or through SigmaAldrich.com/nextgenoligos.
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What cell types are MULTIseq compatible with?
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The MULTIseq Anchor/Co-anchor Lipid Modified Oligos can embed into any lipid bilayer. MULTIseq oligos are cell-type agnostic and can be implanted into any cell membrane. MULTIseq oligos are also compatible with purified nuclei with no changes to the labeling procedure.
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How can I ensure a high cell yield during labeling?
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Start with as healthy cells as possible. Process cells efficiently and store them on ice when feasible. Carefully pipette cells to ensure isolated cells/nuclei are free from clumps and aggregates during labeling/wash steps. Aspirate slowly with a pipette retaining 50-100 µl of supernatant to ensure the pellet is not disrupted. Free floating DNA from dead cells can cause stickiness that could lead to straining/filtration cell loss.
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How will adding MULTIseq to my scRNAseq experiments impact my workflow?
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MULTIseq oligos are simple to use and have minimal impact on the overall scRNAseq process. During the harvest/cell prep, individual samples are labeled with the MULTIseq Anchor/Co-anchor LMOs and a unique MULTIseq barcode. The labeling process will add 30 min to an hour to the overall process. During the SPRI bead cDNA clean-up step, the supernatant is retained and will contain the MULTIseq barcodes. The supernatant will be subjected to its own separate clean-up and library prep.
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What scRNAseq platforms is MULTIseq compatible with?
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The MULTIseq LMOs are compatible with 10X Genomics, BD Rhapsody, Drop-Seq, iCell8, etc.
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Will MULTIseq work with 5’ or 3’ capture chemistry?
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The capture chemistry that will be utilized (5’ vs 3’) will determine the barcodes that are used with the MULTIseq LMOs. We have barcodes for both 5’ or 3’ chemistry. The MULTIseq Anchor/Co-anchor LMOs anchor in cells/nuclei completely independent of the capture chemistry that will be utilized.
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I will be submitting my MULTIseq labeled cells to our core facility for single cell processing and deep sequencing. How will MULTIseq labeled samples impact the workflow?
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First, if you wish to “superload” the lane(s), you will need to specify to have 40K cells loaded per lane instead of the normal 15K. Second, during the 0.6X SPRI Bead cDNA clean-up in Step 2.3, the supernatant MUST be saved. The supernatant is normally discarded but will contain the MULTIseq Barcodes. The barcodes must be cleaned up and library prepped separate from the cDNA. Once the MULTIseq barcode library has been generated, it can be sequenced together with the cDNA library at 2-3K MULTIseq barcode reads per cell.
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How will the raw deep sequencing files (FASTQ) be processed for the gene expression library vs the MULTIseq barcode library?
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The FASTQ files generated from the gene expression library will be processed through the 10X Cell Ranger pipeline, as usual. The MULTIseq barcode library FASTQ files will be processed deMULTIplex2 (https://github.com/Gartner-Lab/deMULTIplex2).
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How many samples can we multiplex together using MULTIseq?
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MULTIseq can multiplex up to 96 separate samples together and are dependent on the unique MULTIseq barcodes that are used. The MULTIseq LMOs are available in 30 and 100 reaction sizes. The MULTIseq barcodes are available with 96 unique sequences.
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