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Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 933 results for: Kin

Showing: 141 - 160 of 933 results

  1. Protocol - How to Streak a Plate

    Type
    Protocol
    ... Protocols Streaking Bacteria on LB Agar Plate Streaking and Isolating Bacteria on an...an LB Agar Plate You may also like... Making LB Agar Plates Bacterial Transformation Recovering Plasmid...explains how to isolate a single bacterial colony by streaking it onto an LB agar plate. Last Update: Feb. 28...down with a paper towel. Maintain sterility by working near a flame or bunsen burner. Obtain the approrpriate...technique is to draw in discontinuous lines. Start by streaking a vertical line of bacteria along one edge of ...

  2. Protocol - Bacterial Transformation

    Type
    Protocol
    ...to isolate single bacterial colonies. Equipment Shaking incubator at 37 °C Stationary incubator at 37 °... microcentrifuge or falcon tube. GENTLY mix by flicking the bottom of the tube with your finger a few ...without antibiotic) to the bacteria and grow in 37°C shaking incubator for 45 min. Pro-Tip This outgrowth step...fast and easy to use, but are less efficient at taking up larger plasmids. If you need to transform large...to ensure that your transformation procedure is working. TIP: Sometimes less is more. Although it may be...

  3. Pouring LB Agar Plates

    Type
    Protocol
    ...Antibiotic Recommended Stock Concentration Recommended Working Concentration Ampicillin 100 mg/mL 100 µg/mL Bleocin...appropriate sterilization technique if you are working with any weird and wonderful organisms. While your...station: Find an empty section of lab bench with a working flame. Spray down the bench with a 70% ethanol ...stop pouring and re-make the gel-mix. If you’re making plates without any antibiotic you can alternatively...viable. You can check for this possibility by streaking out both strains on plates without any antibiotic...

  4. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...Microcentrifuge Electrophoresis chamber Power supply Rocking platform Fume hood Metal spatula Razor blade Plastic...deionized water for 5 min with gentle agitation on a rocking platform. Pour off the water in the sink. Add 20...and incubate for 1 h with gentle agitation on a rocking platform. Pour off the SimplyBlue SafeStain in ...and incubate for 1 h with gentle agitation on a rocking platform. Pour off the water in the sink. Add 100...and incubate for 1 h with gentle agitation on a rocking platform. Pour off the water in the sink. Take ...

  5. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...15,000 human and 15,000 mouse genes. Addgene is working with the TRC to make this shRNA cloning vector ...the transduced cells. hPGK Human phosphoglycerate kinase promoter drives expression of puromycin. Puro R...it has been diluted. Mix by swirling or gently flicking the tube. Incubate for 5 minutes at room temperature...the walls of the tube. Mix by swirling or gently flicking the tube. f. Incubate for 20-30 minutes at room...errors may occur. Addgene makes no warranty of any kind regarding the contents of any literature. Literature...IS” “AS AVAILABLE” basis without warranty of any kind either expressed or implied, including but not limited...

  6. Protocol - How to Perform Sequence Analysis

    Type
    Protocol
    ...same location. Looking at the trace file will give you more information than simply looking at the bases.... You can find Addgene's sequencing results by clicking on the "View Sequences" link on the Plasmid Information...results may indicate bases at specific locations, by looking at the trace file, you will see that these base...

  7. Weighing Reagents Protocol

    Type
    Protocol
    ... or other solutions. A key part of this task is making sure you’re weighing all reagents precisely to ...capacity for the material that you are weighing by looking for a weight range on the scale. Make sure that...how to properly dispose of reagents that you are working with. Pro-Tip When you weigh out a reagent, you...you have your reagents weighed out, you can begin making your solutions!...

  8. Lentivirus Production

    Type
    Protocol
    ...There can be batch to batch variation when making the PEI working stock, therefore the ratio of μg DNA:μg...After 2 months, discard the tube and thaw a new working stock. The optimal mass DNA:mass PEI ratio will...mixture. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate the mixture 12–...

  9. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...add 385µl of water. After making your 100uM stock, immediately make a working concentration of each primer...primer (10uM) by making a 1:10 dilution of the stock. For example, add 100µl of primer stock to 900µl of...cloning purposes. What do I do if my PCR isn't working? Try adding 1µl of 25mM MgCl 2 and/or 1µl DMSO ...

  10. Kit Free RNA Extraction

    Type
    Protocol
    ...or RNase AWAY®, may be used). For more tips on working with RNA, read this blog post on RNA extraction...well-ventilated space and under a fume hood when working with the volatile reagents in the list above. Procedure...freeze-thaw cycles of your entire RNA sample, consider making smaller aliquots of it and storing those in -80...

  11. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...2 Labs Learn how to best protect yourself when working in BSL-1 and BSL-2 labs Watch the Video! Lab Safety...Description (Link opens in a new window) Link to Video Making LB Agar Plates Create plates to culture bacteria...antibiotic to a pre-poured plate Watch the Video! Streaking Bacteria Isolate single bacterial colonies on ...

  12. General Transfection

    Type
    Protocol
    ...After 2 months, discard the tube and thaw a new working stock. Considerations Before You Start The health...DNA. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate the mixture 15–...DMEM complete. Incubate the cells 24–48 h before checking for protein expression. Sample Data Legend: Lenti-X...

  13. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...Automated cell counter 37 °C, 5% CO 2 incubator with shaking platform set to 120 rpm 37 °C bead bath Vortex ...Millipore Sigma A6279 Before Starting Warm the DNA and working stock of PEI-MAX to room temperature before use...flask.Incubate in a 37 °C, 5% CO 2 incubator on a shaking platform set to 120 rpm. Pro-Tip Do not use cells...

  14. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...In its simplest form, PCR based cloning is about making a copy of a piece of DNA and at the same time adding...restriction site (GAATTC) to the 5’ end of this primer, making our Forward Primer 5'-GAATTCATGTGGCATATCTCGAAGTAC...get PCR amplification. We can start similarly, taking the final 18bases of the ORF, including the stop...

  15. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...Workflow Timeline Day 1: Antigen Coating Day 2: Blocking Day 3: Primary antibody incubation Day 4: Secondary...aspirate the wash buffer from the wells. Prepare the blocking buffer (1% BSA in PBS) as follows: Add 250 mg ...mix. Using a multichannel pipette, add 200 µL of blocking buffer to each well. Cover the plate with a plate...

  17. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...sites, will result in similar sized bands, thus making this simple digest less informative. This is particularly...freezer and you are not 100% sure it is what you are looking for, but you have a map and know exactly what it...

  18. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...Liquid Bacterial Culture You may also like... Streaking and Isolating Bacteria on an LB Agar Plate Creating...Incubate bacterial culture at 37°C for 12-18 hr in a shaking incubator. Note: Some plasmids or strains require...

  19. Centrifugation

    Type
    Protocol
    ...appropriate for the lab space in which you are working. Even if your samples may not require specific ...centrifuge is clean and that everything appears to be working smoothly. Place your sample tubes into the rotor...

  20. Gibson Assembly Protocol

    Type
    Protocol
    ... with a two-part Gibson reaction if you're only making a small change in a plasmid (such as point mutations... fine in an assembly if you want to save time. Working on ice, combine segments in the Gibson Assembly...
Showing: 141 - 160 of 933 results