Product info: GLAD-PCR assay Kit


Name
GLAD-PCR assay Kit
Cat. #K009SK009L
Package, reactions2001000

DescriptionGLAD-PCR assay Kit was developed for 200 and 1000 GLAD-PCR reactions.
The following reagents are supplied with this product K009S:
1. 1x TE Buffer - 200 μL,
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 (200 u.a./μL) - 100 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 - 25 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

The following reagents are supplied with this product K009L:
1. 1x TE Buffer - 1 mL,
2. 10x SE TMN Buffer - 750 μL,
3. DMSO - 175 μL,
4. BSA, 10 mg/mL - 350 μL,
5. GlaI (20 u.a./μL) - 35 μL,
6. Universal adapter, double-stranded (10 μM) - 550 μL,
7. ATP, 10 mM - 550 μL,
8. DNA ligase T4 (200 u.a./μL) - 450 μL,
9. 10X SE GLAD Buffer - 2200 μL,
10. MgCl2, 50 mM - 600 μL,
11. dNTP Mix, 10 mM each - 450 μL,
12. SP Taq DNA Polymerase, 5 u.a./μL - 200 μL,
13. Control DNA Raji, 18 ng/μL - 50 μL,
14. Control DNA HeLa, 18 ng/μL - 50 μL,
15. Control DNA Lambda, 18 ng/μL - 125 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

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.
Application- Study of DNA methylation
- Determination of R(5mC)GY site in the point of interest in human DNA
Storage conditionsStore at -20°C.
NotesReaction requires:
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 (see Cat.# K010/1, K010/3, K010/5 and K010/32).
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 (see Instruction manual).
MSDS:Download MSDS as PDF
References:1. A.G. Akishev et al. "GLAD-PCR assay of selected R(5mC)GY sites in URB1 and CEPBD genes in human genome" // Res J Pharm Biol Chem Sci, 8(1), 2017
2.
GLAD-PCR Assay of DNA Methylation Markers Associated with Colorectal Cancer // Evdokimov et al.,Biol Med (Aligarh) 2016, 8:7
3.GLAD PCR Assay Kit instruction manual
4.GLAD PCR Assay demonstration on control DNAs