GLAD-PCR assay Kit was developed for 200 GLAD-PCR reactions. The following reagents are supplied with this product:
1. dH2O -4 mL,
2. 10x SE TMN Buffer - 150 μL,
3. DMSO - 35 μL,
4. BSA, 10 mg/mL - 70 μL,
5. GlaI (20 u.a./μL) - 10 μL,
6. Universal adapter, double-stranded (10 μM) - 110 μL,
7. ATP, 10 mM - 110 μL,
8. DNA ligase T4 concentrated (2000 u.a./μL) - 12 μL,
9. 10X SE GLAD Buffer - 430 μL,
10. MgCl2, 50 mM - 120 μL,
11. dNTP Mix, 10 mM each - 90 μL,
12. SP Taq DNA Polymerase, 5 u.a./μL - 40 μL,
13. Control DNA Raji, 18 ng/μL - 10 μL,
14. Control DNA HeLa, 18 ng/μL - 10 μL,
15. Control DNA Lambda, 18 ng/μL - 10 μL,
16. Control URB1 mix (oligonucleotide primers + TaqMan probe, 10 μM each) - 40 μL,
17. Control CEBPD mix (oligonucleotide primers + TaqMan probe, 10 μM each) - 15 μL
18. Hybrid primers1 (10 μM) - 170 μL.
1 - You can choose 1 (Cat.# K010/1), 3 (Cat.# K010/3), 5 (Cat.# K010/5) of hybrid primers, or order full range of 32 hybrid primers (Cat.# K010/1). The list of 32 hybrid primers (HP1-HP32) you can find in the Instruction manual
Additional services are available with GLAD-PCR assay Kit order:
1. Primers and TaqMan probes design (free for up to 3 RCGY sites);
2. Primes and TaqMan probes synthesis - price upon request;
3. GLAD PCR assay with synthesized oligonucleotides on selected DNA from malignant cell lines (see SE list of products) - price upon request.
- Study of DNA methylation - Determination of R(5mC)GY site in the point of interest in human DNA
Store at -20°C.
1. DNA sample,
2. Genome primer and TaqMan probe designed for DNA region of interest,
3. Hybrid primer (includes constant part complimentary to universal adapter and tetranucleotide part which is complimentary to DNA at the point of GlaI hydrolysis). Hybrid primers may be ordered by request. The Kits K010/1, K010/3, K010/5 and K010/32 includes 1, 3, 5 of chosen primers, or full range of 32 hybrid primers, respectively. The concentration of each hybrid primer is 10 μM, quantity - 170 μL.
All reagents for the control experiments are included.
The time of assay including DNA hydrolysis, ligation of the adaptor and PCR is less than 4 hours.