Abstract
SV-40 DNA sheared form was coupled in a stable covalent bond to cyanogen bromide activated Sepharose. Under the conditions used at least 80% of the DNA was bound to Sepharose. The T1/2 of hybridization of 0.5 μg/ml of SV-40 cRNA to SV-40 DNA-Sepharose was 1 hr. This rate of hybridization is sufficiently rapid to purify SV-40 sequences from solutions containing as little as 0.05-0.1 μg/ml. Nonspecific hybridization of RNA is in the range of 0.1-0.2% of the total input RNA. The DNA-Sepharose is fairly stable and can be reused several times to purify RNA. The SV-40 DNA-Sepharose was used to select large quantities of virus specific RNA from SV-40 infected BS-C-1 cells. The virus specific RNA when added to cell-free extracts from wheat germ was shown to direct the synthesis of the major viral structural protein VP-1.
| Original language | English |
|---|---|
| Pages (from-to) | 4215-4220 |
| Number of pages | 6 |
| Journal | Biochemistry |
| Volume | 14 |
| Issue number | 19 |
| State | Published - Dec 1 1975 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biochemistry
Cite this
Purification of SV-40 messenger RNA by hybridization to SV-40 DNA covalently bound to sepharose. / Gilboa, Eli; Prives, Carol L.; Aviv, Haim.
In: Biochemistry, Vol. 14, No. 19, 01.12.1975, p. 4215-4220.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Purification of SV-40 messenger RNA by hybridization to SV-40 DNA covalently bound to sepharose
AU - Gilboa, Eli
AU - Prives, Carol L.
AU - Aviv, Haim
PY - 1975/12/1
Y1 - 1975/12/1
N2 - SV-40 DNA sheared form was coupled in a stable covalent bond to cyanogen bromide activated Sepharose. Under the conditions used at least 80% of the DNA was bound to Sepharose. The T1/2 of hybridization of 0.5 μg/ml of SV-40 cRNA to SV-40 DNA-Sepharose was 1 hr. This rate of hybridization is sufficiently rapid to purify SV-40 sequences from solutions containing as little as 0.05-0.1 μg/ml. Nonspecific hybridization of RNA is in the range of 0.1-0.2% of the total input RNA. The DNA-Sepharose is fairly stable and can be reused several times to purify RNA. The SV-40 DNA-Sepharose was used to select large quantities of virus specific RNA from SV-40 infected BS-C-1 cells. The virus specific RNA when added to cell-free extracts from wheat germ was shown to direct the synthesis of the major viral structural protein VP-1.
AB - SV-40 DNA sheared form was coupled in a stable covalent bond to cyanogen bromide activated Sepharose. Under the conditions used at least 80% of the DNA was bound to Sepharose. The T1/2 of hybridization of 0.5 μg/ml of SV-40 cRNA to SV-40 DNA-Sepharose was 1 hr. This rate of hybridization is sufficiently rapid to purify SV-40 sequences from solutions containing as little as 0.05-0.1 μg/ml. Nonspecific hybridization of RNA is in the range of 0.1-0.2% of the total input RNA. The DNA-Sepharose is fairly stable and can be reused several times to purify RNA. The SV-40 DNA-Sepharose was used to select large quantities of virus specific RNA from SV-40 infected BS-C-1 cells. The virus specific RNA when added to cell-free extracts from wheat germ was shown to direct the synthesis of the major viral structural protein VP-1.
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UR - http://www.scopus.com/inward/citedby.url?scp=0016752161&partnerID=8YFLogxK
M3 - Article
C2 - 170957
AN - SCOPUS:0016752161
VL - 14
SP - 4215
EP - 4220
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 19
ER -