High-throughput BCR Sequencing of Sorted Cells
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : Harvard University
Other organizations : Icahn School of Medicine at Mount Sinai
Variable analysis
- Whole transcriptome amplification (WTA) on the sorted cell-lysates according to the Smart-Seq2 protocol
- Amplification of heavy and light chain sequences from the WTA products utilizing pools of partially degenerate pools of V region specific primers (Qiagen HotStar Taq Plus)
- Addition of cellular barcodes and index adapters (based on Nextera XT Index Adapters, Illumina Inc.) using a step-out PCR method
- Heavy and light chain sequences obtained after demultiplexing, pairing, and overlapping the sequence reads (Panda-Seq) and aligning against the human IMGT database
- Not explicitly mentioned
- Positive control: None mentioned
- Negative control: None mentioned
Annotations
Based on most similar protocols
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!