muriana at aclcb.purdue.edu
Thu Dec 19 11:49:11 EST 1991
In article <1991Dec18.224447.8529 at casbah.acns.nwu.edu>, rcbrown at casbah.acns.nwu.edu (Robert Brown) writes:
>In article <1991Dec18.213347.27747 at magnus.acs.ohio-state.edu> cbeall at magnus.acs.ohio-state.edu (Clifford J Beall) writes:
>>Dear Methods people,
>>Does anyone have a method for subtracting cDNA prior to PCR?
>>We are using degenerate primers and want to clone a cell specific
>>message. It seems one could hybridize with [+] cDNA with [-] RNA,
>>remove hybrids, then treat the remainder with RNAse. Is HAP still the
>>method of choice for hybrid removal and does anyone know conditions
>>for this? Thanks in advance.
>> Cliff Beall
>> Ohio State Biotech. Ctr.
>Invitrogen sells a kit for subtractive hybridization using biotinylation of
>the [-] RNA and extraction of the hybrid into phenol after binding
Here are some references regarding cDNA subtraction:
Nucleic Acid Res, 14:10027, 1986 ; Welcher, Torres, & Ward
"Selective enrichment of specific DNA, cDNA, and RNA
sequences using biotinylated probes, avidin, and
Nucleic Acids Res, 17:807, 1989; Kuze, Shimizu, and Honjo
"A new vector and RNase H method for the subtractive hybridization"
Nucleic Acids Res, 18:4833, 1990; Rubenstein et al.
"Subtractive hybridization system using single-stranded phagemids
with directional inserts"
Anal. Biochem, 187:364-373, 1990; Fargnoli, Holbrook, and Fornace
"Low-ratio hybridization subtraction"
PNAS, 87:2720-2724, 1990; Wieland, Bolger, Asouline and Wigler
"A method for difference cloning: gene amplification following
Anal. Biochem., 186:369-373, 1990; Barr and Emanuel
"Application of a subtraction hybridization technique involving
photoactivatable biotin and organic extraction to solution
hybridization analysis of genomic DNA"
Nucleic Acids Res, 19:1954, 1991; Swaroop et al.
"A simple and efficient cDNA library subtraction procedure:
isolation of human retina-specific cDNA clones"
Hope this helps,
Internet: muriana at aclcb.purdue.edu
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