How long can I store the cDNA from my reverse transcription step?
You can store your cDNA at 2-6 degrees C for up to 24 hours. For long-term storage, store the cDNA at -15 to -25 degrees C and add EDTA to a final concentration of 1 mM to prevent degradation.
I'd like to generate double-stranded DNA (dsDNA) from mRNA. Do you have a kit for this?
1.SuperScript Plasmid System for cDNA Synthesis and Cloning
-Generating and cloning cDNA into a CMV-containing Gateway-compatible vector (pCMVSport6) for mammalian screening
-mRNA required as starting material (must have poly(A) tail)
-Can probe screen in E.coli but not expression screen
-Resulting dsDNA is unidirectional with Sal I / Not I sites
2.SuperScript Choice System for cDNA Synthesis
-Will work on any RNA source
-Can use random, oligo(dT), or combination
-Will result in cDNA with EcoRI ends which can be cloned into any vector (plasmid or phage) that has EcoRI sites
-Good for generation of cDNA libraries
-Resulting dsDNA will clone bidirectionally
3.SuperScript Double-Stranded cDNA Synthesis Kit
-For generating blunt-ended, double-stranded cDNA
-Will work for total RNA or mRNA as starting material
引用和文献 (1)
引用和文献
Abstract
Gene expression comparison of biopsies from Duchenne muscular dystrophy (DMD) and normal skeletal muscle.
Authors: Haslett Judith N.; Sanoudou Despina; Kho Alvin T.; Bennett Richard R.; Greenberg Steven A.; Kohane Isaac S.; Beggs Alan H.; Kunkel Louis M.;
Journal:Proc Natl Acad Sci U S A
PubMed ID:12415109
'The primary cause of Duchenne muscular dystrophy (DMD) is a mutation in the dystrophin gene leading to the absence of the corresponding RNA transcript and protein. Absence of dystrophin leads to disruption of the dystrophin-associated protein complex and substantial changes in skeletal muscle pathology. Although the histological pathology of dystrophic ... More