Ultraviolet stress-related gene of highland barley and application thereof
A highland barley and gene technology is applied to the ultraviolet stress-related genes of highland barley and its application field, which can solve the problems of affecting the growth and development of highland barley, no effective means, and yield impact, and achieve the effect of good market application prospects.
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Embodiment 1
[0031] Cloning and expression of embodiment 1 gene of the present invention
[0033] (1) RNA extraction and reverse transcription
[0034] 1. Use highland barley 320 and hi 8 seeds to germinate and grow into seedlings.
[0035] 2. Take fresh leaves and use plant extraction kit to extract RNA and detect.
[0036] 3. The extraction steps are as follows:
[0037] 3.1 Instruments and reagents
[0038] Main instruments: SCILOGEX D3024R centrifuge, analytikjena-Easycycler; Scientz-48 high-throughput tissue grinder;
[0039] Main reagents: Reverse Transcription Kit (TUREscript 1st Stand cDNA SYNTHESIS Kit) (Aidelai, Cat. No. PC1802), 2× master mix (Germany DBI pcrMix).
[0040] 3.2 Experimental methods and steps
[0041] 3.2.1 Extraction of RNA from tissue samples
[0042] The RNA extraction kit uses Jena innuPREP RNA Mini Kit.
[0043] 3.2.2 cDNA synthesis (reverse transcription)
[0044] Use Aidlab’s reverse transcription kit (TUREscript 1st Stand ...
Embodiment 2
[0116] Embodiment 2 The correlation between the gene of the present invention and the highland barley being subjected to ultraviolet radiation stress
[0117] 1. Materials
[0118] Two seedlings of highland barley have been provided, and the sample information is as follows:
[0119]
[0120] 2. Instruments and reagents
[0121] 2.1 Main Instruments
[0122] analytikjena-qTOWER2.2 fluorescent quantitative PCR instrument (Germany), SCILOGEX D3024R centrifuge (USA), general PCR instrument analytikjena-Easycycler (Germany); ultra-micro nucleic acid protein analyzer scandrop100 (Germany); pipette (USA bio- rad), DNA electrophoresis spectrum viewer (Junyi), electrophoresis instrument (Junyi).
[0123] 2.2 Main reagent consumables
[0124] Reverse transcription kit (TUREscript 1st Stand cDNA SYNTHESIS Kit) (Aidelai (is a brand) article number PC1802,); 2× Green master mix (Germany DBI). 10ul tip (US GCS), 200ul tip (US GCS), 1ml tip (US GCS), 200ul RNase-free PCR reaction tu...
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