Skip to main content
Addgene

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

Information

In Category

We narrowed to 48 results for: IND;

Showing: 11 - 20 of 48 results

  1. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ...sticky ends", are what allow the vector and insert to bind to each other. When the sticky ends are compatible...standard ligation reaction. Use a (Link opens in a new window) ligation calculator to easily quantify how much...take a fresh tube, thaw it one time and aliquot individual tubes of 5, 10 or 20μL for storage at -20°C. ...troubleshooting failed ligations. The following table indicates the various controls: Control Ligase Interpretation... treated vector Insert or water + Any colonies indicate contamination of intact plasmid in ligation or...every available vector end. (Link opens in a new window) Ligation calculators are easily found on the web...exactly how much of each to use. Reference Page Top Index...

  2. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...that the only version of full-length cDNA you can find for YGOI is in a bacterial expression vector. Using...are creating a fusion protein) Ideally, you will find two different restriction enzymes for your subcloning...cloned in the correct orientation. If you cannot find enzymes that meet these criteria, do not fear. You...restriction enzyme buffer (see (Link opens in a new window) New England Biolabs for more information about...from your transformation will give you the first indication as to whether your transformation worked. our...Finished Plasmid Finally, you will need to pick individual bacterial colonies and check them for successful... and can begin your studies. Reference Page Top Index...

  3. CRISPR Library Amplification

    Type
    Protocol
    ...recombination can all impact the representation of individual plasmids in the pooled library during amplification...designed to be as general as possible but note that individual libraries may require modifications dictated ...satisfactory results for libraries up to 200,000 individual plasmids and can be scaled according to the actual...cells is sufficient for libraries up to 200,000 individual plasmids. For larger libraries scale the number...but this dramatically increases the chances of individual plasmids being lost from the pool and/or increasing...like Glycerol or DMSO solutions. Quantify the individual Maxipreps by Nanodrop, Picogreen, or Qubit. Each...or reagents are significantly overloaded. If you find that your pellet outstrips the capacity listed in...

  4. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ... Test a variety of brands and FBS lot numbers to find one suitable with your protocols. FBS can be purchased...Check the cells for mycoplasma (Link opens in a new window) regularly Do not over or under-grow your cells...of resistant cells start proliferating and the individual wells become confluent, expand into larger vessels...Generation of monoclonal cell lines from expansion of individual A549 cells stably expressing Cas9. A549 cells...for 9 days. Single cells were then plated in individual wells of a 96-well plate and left undisturbed...Methods. 2014 Aug;11(8):783-4 (Link opens in a new window) . Last reviewed on: September 9, 2023...

  5. Pouring LB Agar Plates

    Type
    Protocol
    ...can hold 5-10 mL of agar and on which you can individually distinguish a maximum of ~100 colonies Pro-Tip...Tetracycline 10 mg/mL 10 µg/mL Notes: Unless otherwise indicated, the antibiotic powder can be dissolved in dH ..., you should prepare your plate pouring station: Find an empty section of lab bench with a working flame... looks like in your particular plate. For the remainder of the plates, pour directly from the bottle. ...test results. Sample Data In all cases below (-) indicates that the tested strain is not supposed to be resistant... resistant to the antibiotic, (+) indicates that the tested strain is supposed to be resistant to the ...

  6. Colony Formation Titering Assay

    Type
    Protocol
    ...transfection. Test a variety of brands and lots of FBS to find one suitable with your protocols. FBS can be purchased...Check the cells for mycoplasma (Link opens in a new window) regularly Do not over or under-grow your cells... the minimum concentration (Link opens in a new window) required to kill all of the cells. Use this dose...Figure 1: A549 cells were transduced with the indicated serial dilutions of the lentiviral vector pRosetta...Figure 2: A549 cells were transduced with the indicated serial dilutions of the lentiviral vector pHAGE...

  7. Video Library

    Type
    Protocol
    ...Videos Browse Addgene's collection of video content. Find video protocols, tips on using Addgene's resources... real Addgenies! Protocols (Link opens in a new window) Video Link Description Related Page Making LB ...Gel Protocol How-To Videos (Link opens in a new window) Video Link Description Related Page MTA Guide ...Depositing Page Career Videos (Link opens in a new window) Video Link Description Jessica Welch, PhD In this...

  8. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ... The DNA polymerase is also stable enough to now bind to the primer DNA sequence. Extend DNA for 1 minute... to design primers . Note: (Link opens in a new window) Primer3 is an excellent resource for choosing ...Multiply the volume of each reagent by the number of individual PCRs you wish to perform and add 10% extra to... DNA amplification reaction. Reference Page Top Index...

  9. DNA Quantification

    Type
    Protocol
    ... to protein (260/280) is generally used as an indicator of the purity of DNA samples. These days, many...1.80-2.00). Repeat for each sample. Notes: Keep in mind that despite the accuracy of the NanoDrop, if two... DNA in a buffered solution. Reference Page Top Index...

  10. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ...Carbenicillin plated over-agar Plate shows several individual colonies and effective selection. 150 µL of 2...Carbenicillin plated over-agar Plate shows less individual colonies than the 1 mg/mL plate and effective...Carbenicillin plated over-agar Plate shows several individual colonies with smaller size than the 1 mg/mL and...
Showing: 11 - 20 of 48 results