Skip to main content
Addgene

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

In Category

We narrowed to 164 results for: Nut

Showing: 141 - 160 of 164 results

  7. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...after the introduction of auxin range from 10-20 minutes, making AID an appealing tool for studying essential...

  8. Kit Free RNA Extraction

    Type
    Protocol
    ...Incubate sample(s) for 15 minutes on ice and centrifuge the sample(s) for 15 minutes at 12,000 × g at 4°C ...Incubate the sample(s) for 2-3 minutes on ice and centrifuge for 15 minutes at 12,000 × g at 4°C to separate...Allow sample(s) to sit at room temperature for 5 minutes to allow for dissociation of the nucleoprotein ...incubating at -80°C overnight. Centrifuge for 20 minutes at 10,000 x g at 4°C and discard the supernatant...and vortex for a few seconds. Centrifuge for 5 minutes at 10,000 x g at 4 °C and remove the supernatant...disturbing the pellet. Air-dry the pellet for 5-10 minutes. Critical It is important to not let the pellet...

  9. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...DNA for 2 minutes at 72°C. Repeat steps 8-10 for 25-30 cycles. Final Extension for 5 minutes at 72°C. .... Basic PCR Program Initial Denaturation for 2 minutes at 94°C: This initiation step heats the double ...bind to the primer DNA sequence. Extend DNA for 1 minute at 72°C: The Taq polymerase has an optimal temperature...Repeat steps 2-4 25-30 times. Final Extension for 5 minutes at 72°C: A final extension to fill-in any protruding...melting temperature (Tm). Set extension step at 1-2 minutes per kilobase of product depending on whether you...time and temperatures. Initial Denaturation for 2 minutes at 94°C. Denature for 30 seconds at 94°C. Anneal...

  10. Ligation Independent Cloning

    Type
    Protocol
    ...room temperature for 30 minutes. Inactivate T4 Pol by heating to 75° for 20 minutes. Volume (μl) Reagent ... for 5 minutes at room temperature, then add 1 μl of 25 mM EDTA, followed by another 5 minutes at room...reaction may be stopped by heating to 75° for 20 minutes. Step 6: Anneal and Transform Mix your treated ...

  11. CRISPR Library Amplification

    Type
    Protocol
    ...12 mL recovery media at 37 °C (for at least 15 minutes). Prewarm 3X LB Agar + Antibiotic plates at 37 ...tubes of electrocompetent cells on ice for 15-20 minutes or until completely thawed. Chill a box of 200 ...been absorbed by the agar. This usually takes 1-2 minutes. Critical Be careful not to rip or shred the agar...LB-bacteria per plate. Centrifuge tubes (4000 G, 4 ℃, 15 minutes) to pellet bacteria. Decant LB and weigh pellet... plasmids are much smaller and are a smaller nutritional burden on the bacteria. Bacteria that contain...

  12. Plasmid Modification by Annealed Oligo Cloning (with Protocols)

    Type
    Protocol
    ... to start at 95°C for 2 minutes. Then, gradually cool to 25°C over 45 minutes. Ligation Dilute 5μL of ...Place tube in 90-95°C hot block and leave for 3-5 minutes. Remove the hot block from the heat source (turn...allowing for slow cooling to room temperature (~45 minutes). Method #2 Place mixed oligos in a PCR tube. Place...

  13. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ... 10x NEB buffer 2 35 μL ddH 2 O Incubate for 4 minutes at 95°C in a PCR machine or in a beaker of boiling...PCR machine, incubate the sample at 70°C for 10 minutes then slowly cool to room temperature over the period...swirling or gently flicking the tube. Incubate for 5 minutes at room temperature. e. Add 80 μL of FuGENE® master...gently flicking the tube. f. Incubate for 20-30 minutes at room temperature. g. Retrieve HEK-293T cells...media from Day 4. Spin media at 1,250 rpm for 5 minutes to pellet any HEK-293T cells that were inadvertently...

  14. AAV Production in HEK293 Cells

    Type
    Protocol
    ...inactivated in the lab by heating to 56 °C for 30 minutes. 0.45 μm polyethersulfone (PES) filter system, ...and add 35 mL of 0.05% Trypsin/EDTA. Wait ~2-3 minutes for cells to detach. Gently tap the sides of the... and sonicate 5 x 1 sec pulses with at least 5 minutes on ice between each pulse, 50% amplitude. Return...

  15. Pouring LB Agar Plates

    Type
    Protocol
    ...presence of the antibiotic). Luria broth (LB) is a nutrient-rich media commonly used to culture bacteria in... are unable to digest the agar but can gather nutrition from the LB within. The addition of an antibiotic...

  17. Weighing Reagents Protocol

    Type
    Protocol
    ...the balance to zero. Pro-Tip If you’re weighing a minute amount of reagent that will be resuspended in a...

  18. Water Bath Protocol

    Type
    Protocol
    ...heat up quicker. Pro-Tip Set up the water bath 30 minutes to 1 hour before you need to use it to allow the...

  20. Centrifugation

    Type
    Protocol
    ...Relative Centrifugal Force (RCF) or Revolutions Per Minute (RPM), these are not the same units and are not...
Showing: 141 - 160 of 164 results