KAS-seq protocol was applied to 1,000, 5,000, and 10,000 HEK293T cells with the following changes. gDNA was isolated from denoted numbers of N3-kethoxal-labeled cells by using Quick gDNA mini plus kit (Zymo, D4068). After biotinylation, gDNA was fragmented by Tn5 transposase (Illumina, 10527865, 1.5 μL for 1,000 cells, 2 μL for 5,000 cells, 5 μL for 10,000 cells) in a 50 μL volume at 37 ˚C for 30 min, followed by a clean-up by DNA Clean & Concentrator-5 kit (Zymo, D4013). After immunoprecipitation using 5 μL pre-washed Dynabeads MyOne Streptavidin C1, DNA-conjugated beads and corresponding inputs were directed used for library PCR by using i5 and i7 index primers (Illumina, 20027213) and NEBNext Ultra II Q5 Master Mix (NEB, M0544S). The PCR reactions were heated at 5 min at 72 ˚C followed by 10 min at 95 ˚C, and were then amplified by 15 cycles (10 sec at 98 ˚C, 30 sec at 60 ˚C, 1 min at 72 ˚C). The libraries were then cleaned-up by using MinElute PCR purification kit (Qiagen, 28804). A detailed step-by-step low-input KAS-seq protocol is included in the Supplementary Protocol and Protocol Exchange 43 .
Dynabeads myone streptavidin c1
Manufactured by Illumina
Dynabeads MyOne Streptavidin C1 are superparamagnetic beads coated with streptavidin, a protein that binds strongly to biotin. They are designed for efficient and versatile capture and separation of biotinylated molecules.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using dynabeads myone streptavidin c1
1
KAS-seq: Scalable Chromatin Profiling
2
KAS-seq: Scalable Chromatin Profiling
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KAS-seq protocol was applied to 1,000, 5,000, and 10,000 HEK293T cells with the following changes. gDNA was isolated from denoted numbers of N3-kethoxal-labeled cells by using Quick gDNA mini plus kit (Zymo, D4068). After biotinylation, gDNA was fragmented by Tn5 transposase (Illumina, 10527865, 1.5 μL for 1,000 cells, 2 μL for 5,000 cells, 5 μL for 10,000 cells) in a 50 μL volume at 37 ˚C for 30 min, followed by a clean-up by DNA Clean & Concentrator-5 kit (Zymo, D4013). After immunoprecipitation using 5 μL pre-washed Dynabeads MyOne Streptavidin C1, DNA-conjugated beads and corresponding inputs were directed used for library PCR by using i5 and i7 index primers (Illumina, 20027213) and NEBNext Ultra II Q5 Master Mix (NEB, M0544S). The PCR reactions were heated at 5 min at 72 ˚C followed by 10 min at 95 ˚C, and were then amplified by 15 cycles (10 sec at 98 ˚C, 30 sec at 60 ˚C, 1 min at 72 ˚C). The libraries were then cleaned-up by using MinElute PCR purification kit (Qiagen, 28804). A detailed step-by-step low-input KAS-seq protocol is included in the Supplementary Protocol and Protocol Exchange 43 .
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