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09-14-2020, 02:56 AM
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#1 |
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Junior Member
Location: UK Join Date: Sep 2020
Posts: 3
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RNA-seq library prep from ~100cells
Hi all,
I'm entirely new to RNA sequencing and am looking for some advice re: library prep with low cells numbers. I've started a new project and am looking to do bulk RNA seq (Illumina) on different populations of FACS sorted hematopoietic stem and progenitor cells from mouse bone marrow. Some of these populations are estimated to be 50-200 cells per sample. I'm aiming to do the RNA extraction and library preparation myself but there seems to be loads of different kits available for low (or ultra low) RNA input. Would be great to know what people have had success with before! Any advice, experience or recommendations for library prep with ~100 HSPCs would be massively appreciated. Thanks! |
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09-14-2020, 11:23 PM
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#2 |
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Senior Member
Location: US Join Date: Dec 2010
Posts: 453
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What is your goal? Gene-level studies or transcript level?
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09-15-2020, 02:48 AM
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#3 |
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Junior Member
Location: UK Join Date: Sep 2020
Posts: 3
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Thanks luc. I'm looking to do gene-level differential expression analysis between these populations.
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09-16-2020, 01:58 AM
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#4 |
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Jafar Jabbari
Location: Melbourne Join Date: Jan 2013
Posts: 1,238
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Any of following are compatible with cell(s) input with no RNA extraction:
SMART-Seq v4 3’ DE Kit (requires less sequencing) SMART-Seq® v4 PLUS Kit NEBNext Single Cell/Low Input RNA Library Prep Kit |
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09-16-2020, 07:07 AM
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#5 |
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Junior Member
Location: UK Join Date: Sep 2020
Posts: 3
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Thanks nucacidhunter!
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09-18-2020, 12:26 AM
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#6 |
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Senior Member
Location: US Join Date: Dec 2010
Posts: 453
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Add Qiagen UPX to the list.
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