Description
Nanobody-Tn5 is a recombinant Tn5 transposase fused with nanobody which can specific bind with primary antibodies from two different species (mouse and rabbit).
Product Information
|
Catalog No. |
12595ES01/12595ES02/12595ES03 |
|
specifications |
10 μL/ 80 μL/ 100 μL |
Product Components
|
Component No. |
Component name |
12595ES01 |
12595ES02 |
12595ES03 |
|
12595-A |
Anti-mouse Tn5 |
10 μL |
80 μL |
100 μL |
|
12595-B |
Anti-rabbit Tn5 |
10 μL |
80 μL |
100 μL |
|
12595-C |
Assemble buffer |
10 μL |
80 μL |
100 μL |
Shipping and Storage
Stored at -85° ～ -65℃ for one year.
Figures
H3K4me1:
H3K4me3
H3K9me3
H3K27ac
H3K27me3
H3K27me3
Control: Conventional pA/G-Tn5, Cut&Tag Multi Cut&Tag: Nanobody Tn5
Instruction
Take high-throughput sequencing library preparation as an example, be sure to read the instructions carefully before use.
1. Adapter preparation
1) Reference oligo name and sequence of Illumina platform:
Oligo A-1：5'-AATGATACGGCGACCACCGAGATCTACACATAAGGCTCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-3'
Oligo A-2：5'-AATGATACGGCGACCACCGAGATCTACACATATCCTTCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-3'
Oligo A-3：5'-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-3′
Oligo B：5'-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-3′
Oligo C：5'-phos-CTGTCTCTTATACACATCT-3'
2) Dissolve the Oligo A, Oligo B, and Oligo C to 200 μM.
3) The following reaction systems are formulated respectively:
|
component |
Reaction 1 |
|
Oligo B （200 μM） |
10 μL |
|
Oligo C （200 μM） |
10 μL |
|
total |
20 μL |
|
component |
Reaction 2 |
|
Oligo A-1 or A-2 or A-3（200 μM） |
10 μL |
|
Oligo C （200 μM） |
10 μL |
|
total |
20 μL |
4) Mix reaction 1 and 2 together, and briefly centrifuge to bring the solution back to the bottom of the tube. Place in the PCR instrument, set up the following program and run it: hot lid at 105°C,and then heat at 94℃ for 2 min. After the time was up, the reaction procedure was stopped without opening the hot lid.The PCR tube was naturally cooled in the PCR instrument for 2 h, and then placed at 4℃ for 5 min.
5) After the reaction, mix the equal volumes of Reaction 1 and Reaction 2. Named ad Adapter Mix and stored from – 25 to – 15℃.
2. for transposon generation
1) To configure the following reaction system:
|
component |
volume (μL) |
|
Anti-mouse Tn5 or Anti-rabbit Tn5 |
10 |
|
Adapter mix (50 μM) |
0.5 |
|
Assemble Buffer |
0.5 |
2) Reaction conditions: use the pipette to gently blow and mix well. Perform the reaction at 25℃ for 1 h (hot lid off). The products of reaction can be directly applied to the library preparation or stored from -25 to -15℃.
Notes
1. Transposase is temperature-sensitive, so it is recommended to complete the adapter embedding as soon as possible. The generated transposon can be stored at-25~ -15℃, with a validity period of 1 year.
2. This product is used for scientific research purposes only.
3. For your safety and health, please wear laboratory clothes and wear disposable gloves.