Buffer system mainly for stabilizing protein/enzyme and performance
B buffer
0.15ml
1 Tube
Mainly activated systems such as magnesium ions
Positive control template
0.1ml
1 Tube
Mainly the positive plasmid template is used to test the effectiveness of the kit
Positive control primer mix
0.06ml
1 Tube
Mainly the primer combination of the positive control template
Reagent Guide Manua
16T/bags,48T/Box
3 bags
Reagent technology of protein/enzyme system: freeze-dried powder, freeze-dried microspheres
Principle overview
This kit is based on a room temperature and constant temperature nucleic acid rapid amplification technology: at room temperature and constant temperature, the recombinase and primer form the protein/single-stranded nucleotide complex Rec/ssDNA, with the help of auxiliary proteins and single-stranded binding protein SSB , invade the double-stranded DNA template; form a D-loop region at the invasion site, and start scanning the DNA double-strands; after finding the target region complementary to the primer, the Rec/ssDNA complex disintegrates, and the polymerase also binds to The 3′ end of the primer initiates chain extension. This kit relies on the action of exonuclease at 39 ºC, adding specific molecular probes designed based on the template, and using fluorescence monitoring equipment to achieve real-time monitoring of the amplification process of the target fragment.
Primer design
It is recommended to use primers with a length of 30-35 bp. Primers that are too short will affect amplification speed and detection sensitivity; primers are designed to avoid the formation of secondary structures that affect amplification; the amplicon length is recommended to be 150-300 bp, usually no more than 500 bp.
Fluorescent probe design
The probe sequence does not overlap with the specific primer recognition site, is 46-52 nt in length, and the sequence avoids palindromic sequences, internal secondary structures, and continuous repeated bases. The probe has four modification sites: the middle position ≥ 35 nt from the 5′ end is labeled with a dSpacer (tetrahydrofuran, THF) as the recognition site for exonuclease; the upstream of the THF site is labeled with a fluorescent group, and the downstream Label a quenching group, the distance between the two groups is 2-4 nt; THF is ≥15 nt from the 3′ end, and the 3′ end is labeled with a modifying group, such as an amine group, a phosphate group or a C3-Spacer.
Product features and advantages:
This kit has the advantages of high sensitivity, strong specificity,and short reaction time (only 20 minutes), and the reaction groups are in dry powder state, which is easy to operate and easy to store.
It can be applied to various brands of fluorescence quantitative PCR instruments, constant temperature fluorescence amplification instruments and other fluorescence detection equipment.
Document
This kit is based on a room temperature and constant temperature nucleic acid rapid amplification technology: at room temperature and constant temperature, the recombinase and primer form the protein/single-stranded nucleotide complex Rec/ssDNA, with the help of auxiliary proteins and single-stranded binding protein SSB , invade the double-stranded DNA template; form a D-loop region at the invasion site, and start scanning the DNA double-strands; after finding the target region complementary to the primer, the Rec/ssDNA complex disintegrates, and the polymerase also binds to The 3′ end of the primer initiates chain extension. This kit relies on the action of exonuclease at 39 ºC, adding specific molecular probes designed based on the template, and using fluorescence monitoring equipment to achieve real-time monitoring of the amplification process of the target fragment.
MagPure Stool DNA Kit is specially designed for high throughput DNA extraction from stool samples. It can get high purity microbial DNA from stool samples (≤200mg). This kit is based on magnetic beads purification and unique inhibiting factors adsorption technology, no phenol-chloroform extraction or alcohol precipitation. It can adsorb humic acid and other inhibiting factors in the solution efficiently. DNA can be directly used for downstream applications such as PCR, Viral DNA testing, bacterial DNA testing, etc.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from 100-150mg stool samples
Applications
PCR, southern blot and enzyme digestion, etc.
Purification technology
Magnetic beads technology
Process method
Manual or automatic
Sample type
Stool
Sample amount
100-150 mg
Elution volume
Time per run
Principle
This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Lysozyme. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.
Advantages
Fast – several samples can be extracted in 60 minutes (after digestion)
High purity – unique adsorbent can completely remove inhibitory factors
High recovery – DNA can be recovered at the level of PG
Kit Contents
Contents
D636401
D636402
D636404
Purification Times
48 Preps
96 Preps
400 Preps
MagPure Particles N
1.7 ml
3.4 ml
14 ml
2ml Bead Tubes
48
96
400
RNase A
10 mg
20 mg
75 mg
Proteinase K
24 mg
48 mg
180 mg
Protease Dissolve Buffer
3 ml
5 ml
20 ml
Buffer ATL
60 ml
110 ml
300 ml
PVP-10
1.2 g
2.2 g
6 g
Buffer PCI
50 ml
100 ml
300 ml
Buffer MLE
30 ml
60 ml
180 ml
Buffer GW1*
22 ml
44 ml
132 ml
Elution Buffer
20 ml
20 ml
60 ml
Storage and Stability
MagPure Particles, RNase A and Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
Document
MagPure Stool DNA Kit is specially designed for high throughput DNA extraction from stool samples. It can get high purity microbial DNA from stool samples (≤200mg). This kit is based on magnetic beads purification and unique inhibiting factors adsorption technology, no phenol-chloroform extraction or alcohol precipitation. It can adsorb humic acid and other inhibiting factors in the solution efficiently. DNA can be directly used for downstream applications such as PCR, Viral DNA testing, bacterial DNA testing, etc.
