YeaCell™ Single Cell Purification Kit includes reagents for single-cell emulsion purification, cDNA purification magnetic beads, and DNA purification/size selection magnetic beads. The magnetic beads in this kit offer higher sensitivity and superior performance in nucleic acid recovery, delivering high yield, excellent stability, and good uniformity.

Features

High Sensitivity & Superior Performance: Magnetic beads enable highly efficient nucleic acid recovery with exceptional sensitivity and purity.

High Yield & Stability: Provide consistently high nucleic acid yields with excellent stability and batch-to-batch reproducibility.

Uniform Bead Quality: Ensure reliable and uniform performance in every purification process.

Versatile Applications: Suitable for single-cell GEM demulsification and purification, cDNA cleanup, and library purification.

Components

Storage

Box I: Store at 2–8°C

Box II: Store at –25°C ~ –15°C

Stable for 1 year.

Instructions

1. Emulsion Breaking and Purification

Using the YeaCell™ Single Cell 3ʹ RNA-seq Kit (Yeasen Cat#12520) as an example, the following steps are required after reverse transcription of water-in-oil emulsions (GEMs).

1) Add 125 μL of Recovery Agent (Yeasen Cat#12525 or equivalent) to the sample well. Do not pipette up and down or vortex. Incubate at room temperature for 5 min, then carefully remove 125 μL of the lower oil phase.

[Note]: Recovery Agent and Reducing Agent have similar names but are used for different purposes. Be careful not to confuse them.

2) Prepare Dynabeads Cleanup Mix as shown in Table 1. (Remove Dynabeads MyOne SILANE from the refrigerator and equilibrate to room temperature for at least 30 min. Prepare 80% ethanol in advance.)

Table 1. Preparation of Dynabeads Cleanup Mix

3) Vortex the Dynabeads Cleanup Mix thoroughly and add it to the GEM sample. Pipette up and down 10 times to mix.

4) Incubate at room temperature for 10 min, mixing by pipetting every 5 min.

5) Prepare Elution Solution I as shown in Table 2, vortex to mix, and briefly centrifuge.

Table 2. Preparation of Elution Solution I

6) After the 10 min incubation, place the tube on a magnetic rack. Once the liquid is completely clear, carefully remove the supernatant.

7) Keep the PCR tube on the magnetic rack, add 200 μL of freshly prepared 80% ethanol to wash the beads, incubate at room temperature for 30 sec, then carefully remove the supernatant.

8) Repeat step 7 once.

9) Keep the PCR tube on the magnetic rack, open the lid, and air-dry the beads (approximately 1 min).

10) Remove the tube from the magnetic rack and add 35.5 μL of Elution Solution I. Vortex or gently pipette to mix thoroughly.

11) Incubate at room temperature for 5 min. Briefly centrifuge and place the tube back on the magnetic rack. After the solution clears (~3 min), carefully transfer 35 μL of the supernatant to a new PCR tube.

12) Proceed to the cDNA amplification step of the YeaCell™ Single Cell 3ʹ RNA-seq Kit (Yeasen Cat#12520 or equivalent).

2. cDNA Purification

Using the YeaCell™ Single Cell 3ʹ RNA-seq Kit (Yeasen Cat#12520) as an example, cDNA Clean Beads are used to purify and recover cDNA after cDNA amplification.

1) Preparation: Remove cDNA Clean Beads from the refrigerator and equilibrate to room temperature for at least 30 min. Prepare 80% ethanol.

2) Vortex or invert the cDNA Clean Beads to ensure thorough mixing.

3) Add 60 μL of cDNA Clean Beads (0.6×, Beads:cDNA = 0.6:1) to the cDNA product. Mix by pipetting and incubate at room temperature for 5 min.

4) Briefly centrifuge the PCR tube and place it on a magnetic rack to separate the beads from the liquid. After the solution clears (~5 min), carefully remove the supernatant.

5) Keep the PCR tube on the magnetic rack, add 200 μL of freshly prepared 80% ethanol, incubate at room temperature for 30 sec, then carefully remove the supernatant.

6) Repeat step 5 once.

7) Keep the PCR tube on the magnetic rack, open the lid, and air-dry the beads until slight cracking appears (~1 min).

8) Remove the tube from the magnetic rack and add 40.5 μL Buffer EB. Gently pipette to mix thoroughly. Incubate at room temperature for 5 min. Briefly centrifuge and place the tube on the magnetic rack. After the solution clears (~3 min), carefully transfer 40 μL of the supernatant to a new PCR tube.

9) Quantify the library using Qubit. Store at 4°C for up to 72 hours, at –20°C for one week, or proceed immediately to the next library preparation step.

3. DNA Purification

Refer to the YeaCell™ Single Cell 3ʹ RNA-seq Kit (Yeasen Cat#12520) manual for using DNA Clean Beads to purify or size-select the constructed library.

4. cDNA & Library Quality Control

1) The product obtained after cDNA purification can be analyzed using the High Sensitivity DNA Chip on the Agilent Bioanalyzer 2100 to assess cDNA amplification.

2) The quality of the purified or size-selected library can be evaluated by measuring concentration and fragment size distribution.

Documents:

Safety Data Sheet

18600_MSDS_HB251011_EN.PDF

Manuals

18600_Manual_Ver.EN20251011.pdf