A pilot single-nucleus RNA sequencing experiment was undertaken to compare single-cell versus single-nuclear results from a matched sample. The biopsy from a 58-year-old male was collected fresh and divided into 8 segments, evenly distributed to be processed fresh for single cell RNA sequencing as above, and the remainder was flash frozen in liquid nitrogen. The sample was later retrieved from liquid nitrogen and processed on dry ice according to the protocol in88 (link) with a lysis buffer containing: 0.32 mM sucrose (BioShop SUC507.1), 5 mM CaCl2 (VWR, 97062-820), 3 mM MgCl2 (Thermo Fisher AM9530G), 20 mM Tris-HCl pH 7.5 (Thermo Fisher, 15567027), 0.1% TritonX-100 (Sigma Aldrich T8787-50ML), 0.1 mM EDTA pH 8.0 (Thermo Fisher AM9260G), 40 U/ml Protector RNAse inhibitor (Sigma Aldrich 3335399001) in UltraPure DNAse/RNAse-free water (Thermo Fisher 10977015). The nuclei were captured and sequenced using 10X Genomics Single Cell 3’ v3 Reagents as above.
T8787 50ml
Manufactured by Thermo Fisher Scientific
T8787-50ML is a lab equipment product from Thermo Fisher Scientific. It is a container with a volume of 50 milliliters. The core function of this product is to provide a standard measurement and storage solution for laboratory use.
Automatically generated - may contain errors
2 protocols using t8787 50ml
1
Single-Nucleus RNA Sequencing of Frozen Tissue
2
Single-Nucleus RNA Sequencing Pilot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A pilot single-nucleus RNA sequencing experiment was undertaken to compare single cell versus single nuclear results from a matched sample. The biopsy was collected fresh and divided into 8 segments, evenly distributed to be processed fresh for single cell RNA sequencing as above, and the remainder was flash frozen in liquid nitrogen. The sample was later retrieved from liquid nitrogen and processed on dry ice according to the protocol in 74 with a lysis buffer containing: 0.32 mM sucrose (BioShop SUC507.1), 5 mM CaCl2 (VWR, 97062-820), 3 mM MgCl2 (Thermo Fisher AM9530G), 20 mM Tris-HCl pH 7.5 (Thermo Fisher, 15567027), 0.1% TritonX-100 (Sigma Aldrich T8787-50ML), 0.1 mM EDTA pH 8.0 (Thermo Fisher AM9260G), 40 U/ml Protector RNAse inhibitor (Sigma Aldrich 3335399001) in UltraPure DNAse/RNAse-free water (Thermo Fisher 10977015). The nuclei were captured and sequenced using 10X Genomics Single Cell 3' v3
Reagents as above.
Reagents as above.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!