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pLenti-Bi-cistronic

Catalog Number P0270
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Product Overview

Instructions Ⅰ. Plasmid dry powder (stored at minus 20 degrees, shelf life 90 days, transportation at room temperature) 1. After receiving the dry powder plasmid, please centrifuge at 5000rpm for 1min, and then add 20μl of sterile water to dissolve the plasmid; 2. Thaw the corresponding competent on an ice box, take 2 μl of plasmid and add it to 100 μl of competent for 30min; 3. 42 degrees heat shock for 60s, ice bath for 2min, add 900μl of antibiotic-free LB medium, and culture at 180rpm shaking for 45min; 4. Centrifuge at 6000rpm for 3min, leave 100μl of supernatant to resuspend, and then coat it on the corresponding resistant LB plate; 5.After inverting overnight, if there are too many colonies, the plasmid will be diluted for transformation, and if there are no colonies, 10ul plasmid will be taken for transformation; 6. Pick single colonies into the corresponding resistant LB liquid medium overnight shaking culture, and extract the plasmid as needed. Ⅱ. Liquid plasmid (minus 20 degrees storage, shelf life 90 days, ice pack transportation). 1. After receiving the liquid plasmid, please shake the preservation tube so that the liquid plasmid does not adsorb on the tube wall; 2.Take an appropriate amount of plasmid according to the experimental needs, and store the rest in a minus 20 degree freezer. Ⅲ. Glycerol strains (stored at minus 80 degrees, shelf life of 90 days, ice pack transportation). 1. After receiving the glycerol strain, please shake the preservation tube so that the bacterial solution does not adsorb on the tube wall; 2. Use the inoculation loop to mark the line in four areas on the solid plate, and add the corresponding resistance if there is resistance; 3. Inverted culture in constant temperature incubator for 12~16h; 4. Be sure to pick a single colony and inoculate it in the liquid medium, and add the corresponding resistance if there is resistance; 5. Shake the shaker and culture overnight, and culture the strains according to the needs of the experiment.

Product Disclaimer

  1. For Research Use Only
    All products available through the MiaoLing Plasmid Platform are intended for research use only. They are not intended for clinical trials or for use in humans, including oral administration, injection, or therapeutic use in vivo or ex vivo.
  2. Plasmid Sequence and Experimental Performance
    Plasmid products are provided and sold based on the corresponding reference/template plasmid. MiaoLing guarantees the accuracy of the specified gene sequence based on our sequencing results, but does not guarantee downstream experimental performance, including gene expression, transfection efficiency, protein expression, or other experimental outcomes.
  3. Sequence Verification of Certain Vector Products
    For certain vector products, even the original manufacturer may not have fully verified the complete plasmid sequence. If the sequence of the key functional regions has been confirmed to be correct, the product will be considered a correct and usable vector for research purposes.
  4. After-Sales Period for Small-Pack Plasmid Products
    The after-sales service period for small-pack plasmid products is three months from the date of receipt. Customers are advised to transform and process the plasmid promptly after receipt. After the three-month period, a new purchase may be required for further support or replacement.
  5. Research-Related Risks and Limitation of Liability
    Scientific research is inherently exploratory and involves a significant degree of uncertainty. To the maximum extent permitted by applicable law, MiaoLing shall not be liable for any additional economic losses, indirect losses, or other liabilities arising from the use of our products or from research outcomes.
  6. Citation in Chinese Publications
    For publications in Chinese, please cite the product as follows:
    Product names and IDs are provided by the MiaoLing Plasmid Platform.
  7. Citation in English Publications
    For publications in English, please cite the product as follows:
    “Product Name (Product Item Number) was obtained from MiaoLing Plasmid, China.”

Related Publications

No Literature Available

Product Documents

Plasmid Elements

misc_signal 170..320 /note="腺病毒5型包装信号" RSV promoter 488..714 /note="禽类肉瘤病毒增强子/启动子" 5' LTR (truncated) 715..895 /note="截短型5'长末端重复序列(LTR)源自1型人类免疫缺陷病毒" HIV-1 Psi 942..1067 /note="packaging signal of human immunodeficiency virus type 1" RRE 1557..1790 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." cPPT/CTS 2297..2412 /note="1型人类免疫缺陷病毒的中心多嘌呤道和中心终止序列(缺少第一个T)。" SV40 poly(A) signal 2441..2522 /note="SV40 polyA信号" mini CMV complement(2656..2722) PGK promoter 2733..3232 /note="小鼠磷酸甘油酸激酶1启动子" HA 3309..3335 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /translation="YPYDVPDYA" SV40 promoter 3353..3650 /note="SV40增强子和早期启动子" SV40 ori 3520..3655 /note="SV40复制起点" PuroR 3656..4255 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /note="赋予对普鲁霉素的抗性" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" WPRE 4268..4855 /note="土拨鼠肝炎病毒转录后调控元件" Factor Xa site complement(4739..4750) /codon_start=1 /product="Factor Xa recognition and cleavage site" Cleavage site after base 4738" /translation="IEGR" 3' LTR (Delta-U3) 4865..5098 /note="自失活型3'长末端重复序列(LTR),源自1型人类免疫缺陷病毒" SV40 poly(A) signal 5170..5291 /note="SV40 polyA信号" ori complement(5619..6207) /direction=LEFT /note="高拷贝数 ColE1/pMB1/pBR322/pUC 复制起点" NeoR/KanR 7014..7808 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /note="赋予对新霉素、卡那霉素和G418(金霉素)的抗性。" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" M13 ori complement(7856..8235) /direction=LEFT /note="M13 噬菌体复制起点;箭头指示(+)链合成方向" repeat_region join(8273..8292,1..82) /note="人类5型腺病毒的倒置末端重复序列"
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