HiI'm starting to work whit 5' cytidine on plants. I want to know if it's possible to prepare a stock solution to store it at -20C. In that conditions the drug lose the desmetilant capacity?I've wrote ... [阅读全文]
Hey
I am analyzing the methylation status of two regions: one ecompasses 500bp the other is around 800bp. Could I just designed primers at flanking sites and amply these two regions? I found usually o ... [阅读全文]
I am having huge problems with bisulfite PCR. Southern analysis indicates that my DNA is unmethylated. My first bisulfite treatment also indicated that the DNA was unmethylated. Subsequent treatments ... [阅读全文]
Hi
I looked for CpG islands within the promoter of my gene.MethPrimer and CpG plot analysis revealed that in this genes promoter region there were two big CpG islands at locations 280-603 and 693-859. ... [阅读全文]
DNA methylation is an epigenetic event that affects cell function by altering gene expression and refers to the covalent addition of a methyl groupcatalyzed by DNA methyltransferase (DNMT)to the 5-car ... [阅读全文]
This protocol was written by Jean-Pierre Issabased on Adams et al.* Kam-Wing Jair has made some useful shortcuts that work well if you are careful.Here is .This assay can be used to measure activity i ... [阅读全文]
In situ immunodetection of 5-methylcytosine (5-mC) on squashed cells using anti-methylcytosine
Cross-references are to Schwarzacher & Heslop-Harrison 2000. Practical in situ Hybridization. Bios Ox ... [阅读全文]
I'm looking for a methode to demethyle DNA in vivo during more than 5 days . All I've read about 5-aza-CdT is for maximum 5 days. Why ?
Is there another drug and if yes how does it work ?
thanks
Elea ... [阅读全文]
Protocol written by James Herman*
Methylation-specific PCR (MSP) is a simple rapid and inexpensive method to determine the methylation status of CpG islands. This approach allows the determination of ... [阅读全文]
CpG islands can be variable in length. Anyone know by what percentage range they can typically vary? I can't find much of any herlp on Google.
Cheers
Chris
-Chris Harris-
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Hi
Does enzyme digestion before bisulfite treatment make a big difference - i am having a few problems with incomplete conversion.
If so can anyone reccomend me a good 'general' enzyme to use - I know ... [阅读全文]
Hi. I am a new member of this forum. Currently I am trying to look for some novel methylated gene in NPC cell line. What method should I start with? Pls help. Thank you.
-CHOO-
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Hello everyone I'm new here and I was wondering if I could get some assistance.
In my research of the literature I thought the differences between the two methods were based on the proximity of the pr ... [阅读全文]
Hi there
Is anyone know how short of the CpG island can be? let say if the CG sequence of about 30bp is that possible the short CpG sequence be methylated?
Thanks you!
Pine
-pine-
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Quick question as I have read all sorts of views on this before:
How long can bisufite-treated DNA be stored in -80℃ freezer before it degrades. Is there any reason why it should degrade quicker than ... [阅读全文]
Adapted from Frommer et.al.*
1.Dilute DNA (up to 2 mg) into 50 ml with distilled H2O.
2.Add 5.5 ml of 2M NaOH.
3.Incubate at 37℃ for 10 minutes (to create single stranded DNA).
4.Add 30 ml of 10 m ... [阅读全文]
This is the URL to my gene of interest.
I am trying to estimate the promoter region of this gene so that I can do Meth specific PCR on the promoter..the promoter region of this gene has not been posit ... [阅读全文]
Source: Protocol Online Abstract: Modifying DNA using sodium bisulfite to convert unmethylated cytosines to uracils and subsequently detect methylated cytosines using methylation specific PCR (MSP) te ... [阅读全文]
Contributed by Dr. A. Gratchev
Single Nucleotide Primer Extension is a powerful method which can be used for the precise analysis of methylation in a certain position. The procedure is shown on the fi ... [阅读全文]
I am planning to perform sodium bisulphite sequencing. Currently I am trying to design primers of the promoter region of my target genes. However some of these promoters have not been well-characteriz ... [阅读全文]
DNA methylation is an epigenetic event that affects cell function by altering gene expression and refers to the covalent addition of a methyl group catalyzed by DNA methyltransferase (DNMT) to the 5-c ... [阅读全文]
Source: Protocol Online
Abstract: Modifying DNA using sodium bisulfite to convert unmethylated cytosines to uracils and subsequently detect methylated cytosines using methylation specific PCR (MSP) te ... [阅读全文]
The restriction/modification system in bacteria is a small-scale immune system for protection from infection by foreign DNA.
W. Arber and S. Linn (1969)
Plating efficiencies of bacteriophage lambda (l ... [阅读全文]
Adapted from Frommer et.al.*
1.Dilute DNA (up to 2 mg) into 50 ml with distilled H2O.
2.Add 5.5 ml of 2M NaOH.
3.Incubate at 37℃ for 10 minutes (to create single stranded DNA ).
4.Add 30 ml of 10 ... [阅读全文]
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