Nanobody-Tn5 _ 12595ES

SKU: 12595ES01

Size: 10 μL
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Sale price$365.00

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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.

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