Fwd: DD-PCR

WSchick at aol.com WSchick at aol.com
Tue Nov 12 08:35:59 EST 1996


In a message dated 96-11-11 11:54:44 EST, dissyst at primenet.com (Display
Systems Biotech) writes:

 Forwarded message:
 From:	wudc at nic.bmi.ac.cn
 
 Dear everyone
 
 The ones who have experince doing differential display,I want your help!
 
 I used 0.2 micrograms total RNA to reverse transcription,and one-tenth
 of the cDNA was used in DDPCR,but I got very faint bands,I think the
 reason may be too little cDNA was used in the reaction.so I want to
 know,how much total RNA is used to do reverse-transciption ,and how much
 cDNA is used to do DDPCR in your reaction?
 If you have some tips or skills ,let me know.Thank you very much!
 
 
 Answer,
 
 The amount of RNA and following cDNA synthesis is extremly important in
 DDRT-PCR analysis.
 I have now been in the DD-PCR world since the beginning back in 92. 
 If you want to read more about our protocols please read some of
 the most important publications in this field, if you want we can sent
 copies.  In our DD kits all the parameter are completely optimized. 
 
 Best Regards
 
 Peter Warthoe, 
 Display Systems Biotech   http://www.displaysystems.com
 
 Some method papers:
 (please start with the chapter in the new Cold Spring Harbor book page
 421):
 
 Bauer, D., Warthoe, P., and Strauss, M. Identification of differentially
 expressed mRNA species by an improved display technique (DDRT-PCR)
 (1993). NAR 21: 4272-4280
 
 Warthoe, P., et al. Detection and identification of expressed genes by
 differential display (1995).  PCR Primers: A Laboratory Manual. Cold
 Spring Harbor Laboratory Press.
 
 Bauer, D.,  Warthoe, P., Liang, P., Averboukh, L., Rohrwild, M., Muller,
 H., Strauss, M., and Pardee, AB. Analysis of altered gene expression by
 differential display (1995). Methods-Enzymol. 254: 304-321 
 
 Rohde, M., Warthoe, P., Gjetting, T., Lukas, J., Bartek, J., and
 Strauss, M. The retinoblstoma protein modulates expression of genes
 coding for diverse classes of proteins including components of the
 extracellular matrix (1996). Oncogene 12: 2393 - 2401.
  >>


---------------------
Forwarded message:
From:	dissyst at primenet.com (Display Systems Biotech)
To:	WSchick at aol.com
Date: 96-11-11 11:54:44 EST

Dear Walt,

The complete address - Answer to the one asking about DD-PCR 

Forwarded message:
From:	wudc at nic.bmi.ac.cn
Sender:	daemon at net.bio.net
Reply-to:	wudc at nic.bmi.ac.cn
To:	methods at net.bio.net
Date: 96-11-06 10:55:58 EST

Dear everyone

The ones who have experince doing differential display,I want your help!

I used 0.2 micrograms total RNA to reverse transcription,and one-tenth
of the cDNA was used in DDPCR,but I got very faint bands,I think the
reason may be too little cDNA was used in the reaction.so I want to
know,how much total RNA is used to do reverse-transciption ,and how much
cDNA is used to do DDPCR in your reaction?
If you have some tips or skills ,let me know.Thank you very much!


Answer,

The amount of RNA and following cDNA synthesis is extremly important in
DDRT-PCR analysis.
I have now been in the DD-PCR world since the beginning back in 92. 
If you want to read more about our protocols please read some of
the most important publications in this field, if you want we can sent
copies.  In our DD kits all the parameter are completely optimized. 

Best Regards

Peter Warthoe, 
Display Systems Biotech   http://www.displaysystems.com

Some method papers:
(please start with the chapter in the new Cold Spring Harbor book page
421):

Bauer, D., Warthoe, P., and Strauss, M. Identification of differentially
expressed mRNA species by an improved display technique (DDRT-PCR)
(1993). NAR 21: 4272-4280

Warthoe, P., et al. Detection and identification of expressed genes by
differential display (1995).  PCR Primers: A Laboratory Manual. Cold
Spring Harbor Laboratory Press.

Bauer, D.,  Warthoe, P., Liang, P., Averboukh, L., Rohrwild, M., Muller,
H., Strauss, M., and Pardee, AB. Analysis of altered gene expression by
differential display (1995). Methods-Enzymol. 254: 304-321 

Rohde, M., Warthoe, P., Gjetting, T., Lukas, J., Bartek, J., and
Strauss, M. The retinoblstoma protein modulates expression of genes
coding for diverse classes of proteins including components of the
extracellular matrix (1996). Oncogene 12: 2393 - 2401.






More information about the Methods mailing list