[Preservation methods and Precautions]
L Transport at room temperature. Store at 2-8℃ for one year. Avoid freezing avoid freezing!
L Magnetic beads must be balanced at room temperature for at least 30 minutes before use.
L The 80% ethanol needs to be used and mixed, otherwise the recovery efficiency will be affected.
L For length separation, the initial sample volume should be ≥100μL, if insufficient, please use ultrapure water to fill. If the sample volume is too small, the pipetting error will increase, which will affect the accuracy of sorting.
L For your safety and health, please wear lab coat and disposable gloves.
[Reference conditions for Library size sorting]
NGSbead was used to construct DNA library. After purification of 1×NGSbead, and then sorting according to the conditions in Table 1, libraries of different sizes were obtained and analyzed with Agilent 2100 Bioanalyzer.
Table 1. Recommended ratio of magnetic bead library sorting
Average length of inserted fragment (BP)
250
250-350.
350-450.
450-550.
550-650.
650-750.
Volume ratio in the first round
0.80 x
0.70 x
0.60 x
0.55 x
0.50 x
0.45 x
Second round volume ratio
0.20 x
0.20 x
0.20 x
0.15 x
0.15 x
0.15 x
Note: 1.8x Bead: All fragment purification; 1.0x Bead: Primers + small amplicons removal; Sorting of 0.65x -- 0.25x Bead fragments.
[Kit composition]
component
100 times
save
PuriMag NGSbead
5 ml
2-8 ℃
Wash buffer
80% ethanol (user supplied)
The normal temperature
Elution buffer
5 ml
The normal temperature
FIG. 2. Flow chart of magnetic bead two-wheel sorting
[Operation Procedure]
1. Preparation
Remove the beads from the refrigerator and balance at room temperature for at least 30 minutes. Prepare 80% ethanol.
2. Double wheel sorting
1) Please scroll or fully reverse the magnetic beads to ensure the mixing.
2) According to the requirements, add the first round of sorting magnetic beads to the DNA solution by referring to Table 1, and mix with vortex mixing or pipette blowing for 10 times.
(3) Incubate at room temperature for 5min.
4) Centrifuge the centrifuge tube briefly and place it in a magnetic rack. After the solution is clarified (about 5min), carefully transfer the supernatant to a clean centrifuge tube.
Note: when transferring supernatant, do not absorb magnetic beads. Even a small amount of residue will affect the quality of subsequent library.
5) Refer to Table 1 to add the second round of sorting magnetic beads to the upward cleaning.
(6) Eddy mixing or pipette blowing for 10 times to mix, and let stand at room temperature for 5min.
7) Place the centrifuge tube in a magnetic rack and remove the supernatant carefully after the solution is clarified (about 5min).
8) Keep the EP tube in the magnetic shelf, add 200μ L freshly prepared 80% ethanol to rinse the magnetic beads, incubate at room temperature for 30 s, and carefully remove the supernatant.
9) Repeat Step 8.
10) Keep the centrifugal tube in the magnetic frame all the time, open the cover and dry the magnetic beads until they just crack (about 5min).
Note: Remember not to dry the magnetic beads for too long, too much drying will affect the purification effect.
11) Take the centrifugal tube out of the magnetic rack, add an appropriate amount of ddH2O (≥20 μL), vortex oscillation or use pipette gently blow and mix thoroughly.
12) Keep the EP tube in the magnetic frame all the time, carefully absorb the supernatant into the clean EP tube, that is to complete the sorting.
Note: This kit is for scientific research purposes only.
|
Warm tip: the products supplied by Beijing Beike Xincai Technology Co., Ltd. are only used for scientific research, not for human body |
| Item ID |
CAS |
ID |
Pack |
Parameter |
Stock |
Make up |
Price |
| BK2021081229 |
|
BK2021081229 |
|
|
100 |
|
$0 |
