Primers for cloning SPFMV (sweet potato feathery mottle virus) O strain and full length genome sequence of SPVC (sweet potato virus C) and cloning method
A feathery mottled virus and genome sequence technology, applied in the field of bioengineering, can solve the problems that it is difficult to obtain the full-length genome sequence of the virus, the full-length genome sequence of the isolate has not been seen, and the content is low
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Embodiment 1
[0065] Embodiment 1 A kind of method of cloning SPFMV-O strain full-length genome sequence
[0066] 1. Materials and methods
[0067] (1) Tested virus material: the leaves of sweet potato plants infected with SPFMV, SPVC and SPCSV were used as materials. The symptoms were as follows: the plants were short, the leaves were shrunken, yellowed, and the veins were distorted. The total RNA was extracted for subsequent experiments.
[0068] (2) Reagents and kits: UNIQ-10 Column Total RNA Extraction Kit was produced by Shanghai Sangon Bioengineering Company; DNA Gel Recovery Kit was purchased from Axygen Biotechnology (Hangzhou) Co., Ltd.; RNase inhibitor, AMV Reverse transcriptase, LATaq DNA polymerase, and pMD19-T vector were purchased from TaKaRa Company, and other commonly used reagents were of domestic analytical grade.
[0069] (3) Primer design: According to the nucleotide sequence of the full-length genome of SPFMV-O registered in GenBank (accession numbers are AB509454, AB4...
Embodiment 2
[0088] Embodiment 2 A kind of method for cloning sweet potato virus C full-length genome sequence
[0089] 1. Materials and methods
[0090] (1) Tested virus material: the leaves of sweet potato plants infected with SPFMV, SPVC and SPCSV were used as materials. The symptoms were as follows: the plants were short, the leaves were shrunken, yellowed, and the veins were distorted. The total RNA was extracted for subsequent experiments.
[0091] (2) Reagents and kits: UNIQ-10 Column Total RNA Extraction Kit was produced by Shanghai Sangon Bioengineering Company; DNA Gel Recovery Kit was purchased from Axygen Biotechnology (Hangzhou) Co., Ltd.; RNase inhibitor, AMV Reverse transcriptase, LATaq DNA polymerase, and pMD19-T vector were purchased from TaKaRa Company, and other commonly used reagents were of domestic analytical grade.
[0092] (3) Primer design: According to the nucleotide sequence of the full-length genome of SPVC registered in GenBank (accession numbers are JX489166,...
Embodiment 3
[0107] Example 3 A method for simultaneous cloning of sweet potato feather mottle virus O strain and sweet potato virus C full-length genome sequence
[0108] 1. Materials and methods
[0109] (1) Tested virus material: the leaves of sweet potato plants infected with SPFMV, SPVC and SPCSV were used as materials. The symptoms were as follows: the plants were short, the leaves were shrunken, yellowed, and the veins were distorted. The total RNA was extracted for subsequent experiments.
[0110] (2) Reagents and kits: UNIQ-10 Column Total RNA Extraction Kit was produced by Shanghai Sangon Bioengineering Company; DNA Gel Recovery Kit was purchased from Axygen Biotechnology (Hangzhou) Co., Ltd.; RNase inhibitor, AMV Reverse transcriptase, LATaq DNA polymerase, and pMD19-T vector were purchased from TaKaRa Company, and other commonly used reagents were of domestic analytical grade.
[0111] (3) Primer design: according to the nucleotide sequences of the full-length genomes of SPFMV...
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