For thermoligation barcoding, the droplet emulsion was incubated as follows (heated lid set to 105 °C, volume set to 100 μl): 12 cycles of [98 °C for 30 s, 59 °C for 2 min], storage at 15 °C. The emulsion was broken by addition of 125 μl Recovery Agent (10x Genomics #220016), and 125 μl of the pink oil phase was removed by pipetting. The remaining sample was mixed with 200 μl of Dynabead Cleanup Master Mix (per reaction: 182 μl of Cleanup Buffer (10x Genomics #2000088), 8 μl of Dynabeads MyOne Silane (Thermo Fisher Scientific #37002D), 5 μl of Reducing Agent B (10x Genomics #2000087) and 5 μl of nuclease-free water). After 10 min of incubation at room temperature, samples were washed twice with 200 μl of freshly prepared 80% ethanol (Merck #603-002-00-5) and eluted in 40.5 μl of EB Buffer (Qiagen #19086) containing 0.1% Tween (Sigma #P7949-500ML) and 1% v/v Reducing Agent B. Bead clumps were sheared with a 10 μl pipette or needle. 40 μl of the sample was transferred to a fresh tube strip and subjected to a 1.0x cleanup with SPRIselect beads (Beckman Coulter #B23318), eluting in 22 μl of EB Buffer.
Reducing agent b
Manufactured by Merck Group
Reducing Agent B is a chemical reagent used in various laboratory applications. It functions as a reducing agent, facilitating the transfer of electrons in chemical reactions. The core purpose of Reducing Agent B is to enable the reduction of target compounds, a fundamental process in many analytical and synthetic procedures. Further details on its specific applications are not provided to maintain an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Eb buffer, by Qiagen (2 mentions)
Dynabead cleanup master mix, by 10x Genomics (2 mentions)
Cleanup buffer, by 10x Genomics (2 mentions)
Reducing agent b, by 10x Genomics (2 mentions)
Recovery agent, by 10x Genomics (2 mentions)
Dynabeads myone silane, by Thermo Fisher Scientific (1 mentions)
Eb buffer, by Beckman Coulter (1 mentions)
Spriselect beads, by Beckman Coulter (1 mentions)
Dynabeads myone silane, by 10x Genomics (1 mentions)
2 protocols using reducing agent b
1
Thermoligation Barcoding Droplet Emulsion Protocol
2
Emulsion Purification and Cleanup Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The emulsion was broken and purified according to the manufacturer’s instructions (10X Genomics). Briefly, 125 µl Recovery Agent (10X Genomics catalog no. 220016) was added to each sample (post GEM-RT incubation) at room temperature for 2 min. Then 125 µl of the pink oil phase was removed by pipetting. The remaining sample was mixed with 200 µl of Dynabead Cleanup Master Mix (per reaction: 182 µl of Cleanup Buffer (10X Genomics, 2,000,088), 8 µl of Dynabeads MyOne Silane (10X Genomics, 2,000,048), 5 µl of Reducing Agent B (10X Genomics, 2,000,087), and 5 µl of nuclease-free water). After 10 min of incubation at room temperature, samples were washed twice with 300 µl of freshly prepared 80% ethanol (Merck, 603–002-00–5) and eluted in 35.5 µl of EB Buffer (Qiagen, 19,086) containing 0.1% Tween (Sigma, P7949-500ML) and 1% v/v Reducing Agent B. Bead clumps were sheared with a 10-µl pipette or needle. Thirty-five µl of the sample was transferred to a new tube.
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!