After MDA amplification adapted in this kit, the coverage of single cell genome can reach more than 95%, and amplification products sizes are between 2kb-100kb, with the average product size greater than 20 kb, and can be widely used in whole genome and exon sequencing, large copy number variation analysis, microsatellite analysis, qPCR analysis and gene chip analysis.
The Phi 29 DNA polymerase used in this kit is a DNA polymerase cloned from a phage and has strong strand displacement activity, which is capable to achieve melting and replication of DNA chains with the complex structure. In addition, Phi 29 DNA polymerase has strong chain affinity, and one single polymerization process can achieve continuous polymerization and extension of up to 100 kb product. The amplification products are applicable to almost all downstream applications; Phi 29 DNA polymerase also has very strong 3 '-5' exonuclease activity, and its fidelity is 1000 times as that of Taq enzyme, which ensure the high fidelity of DNA synthesis.
Normally, one reaction in Discover-scTM Single Cell and Kit V2 can produce a 30-40 μg genomic DNA of high coverage.
- High sensitivity
Whole genome non-differential amplification can be achieved using single cells or extremely small amounts of DNA as templates.
- High coverage
Coverage of single cell genome can be more than 95% after amplification.
- High uniformity
Based on the non-preference MDA amplification, the whole genome can be amplified uniformly.
- High fidelity
The fidelity of Phi29 DNA polymerase is superior to current most high-fidelity enzymes.
- High yield
6-8 hours of reaction can generate 30-40 ug genomic DNA.
Extensive adaptation of amplified products
The amplification products of Discover-scTM Single Cell Kit V2 are distributed in 2-100kb, with average product size more than 20kb, and can be widely used in whole genome and exon sequencing, large copy number variation analysis, microsatellite analysis, qPCR analysis, and gene chip analysis.
M: DL15000
C: amplification products of single cell genome
D: amplification products of purified genome
High coverage of amplified products! High average!
Uniformity and coverage analysis results of whole genome amplification products of Discover-scTM Single Cell Kit V2.
Using qPCR primers distributed on 16 different chromosomes to detect the uniformity and coverage of the whole genome amplification products of Discover-sc Single Cell Kit V2, in which the template was 5 ng. The results showed high uniformity and coverage of the amplified products.
Kit Components:
|
Components |
N602-01 (24 rxn) |
N602-02 (96 rxn) |
|
|
Box 1 |
Discover-sc DNA Polymerase |
48 μl |
192 μl |
|
Discover-sc Reaction Buffer |
750 μl |
3 ´ 1 ml |
|
|
Buffer D |
1 ml |
2 ´ 1 ml |
|
|
Buffer N |
1 ml |
2 ´ 1 ml |
|
|
DTT, 1M |
1 ml |
1 ml |
|
|
PBS |
1 ml |
2 ´ 1 ml |
|
|
H2O |
1 ml |
2 ´ 1 ml |
|
|
Discover-sc Primer 1 |
30 μl |
120 μl |
|
|
Discover-sc Primer 2 |
30 μl |
120 μl |
|
|
Discover-sc Primer 3 |
30 μl |
120 μl |
|
|
Discover-sc Primer 4 |
30 μl |
120 μl |
|
|
Discover-sc Primer 5 |
30 μl |
120 μl |
|
|
Discover-sc Primer 6 |
30 μl |
120 μl |
|
|
Discover-sc Primer 7 |
30 μl |
120 μl |
|
|
Discover-sc Primer 8 |
30 μl |
120 μl |
|
|
Discover-sc Primer 9 |
30 μl |
120 μl |
|
|
Discover-sc Primer 10 |
30 μl |
120 μl |
|
|
Discover-sc Primer 11 |
30 μl |
120 μl |
|
|
Discover-sc Primer 12 |
30 μl |
120 μl |
|
|
Discover-sc Primer 13 |
30 μl |
120 μl |
|
|
Discover-sc Primer 14 |
30 μl |
120 μl |
|
|
Discover-sc Primer 15 |
30 μl |
120 μl |
|
|
Discover-sc Primer 16 |
30 μl |
120 μl |
|
|
Box 2 |
Discover-sc AceQ Master Mix |
4 ´ 1.25 ml |
16 ´ 1.25 ml |
|
ROX Dye 1 |
200 μl |
4 ´ 200 μl |
|
|
ROX Dye 2 |
200 μl |
4 ´ 200 μl |
Box1: stored at -20°C, if for long-term storage please store at -70°C;
Box2: stored at -20°C, avoid light.