We narrowed to 933 results for: KIN

...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–...

...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 ...

...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...

...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 ...

...Incubate bacterial culture at 37 °C for 12–18 h in a shaking incubator. Note: Some plasmids or strains require... can be dissolved in dH 2 0. Addgene recommends making 1000X stock solutions and storing aliquots at -... cell density. Resources Additional Protocols Streaking and Isolating Bacteria on an LB Agar Plate Creating...

...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...

...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...

...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...

...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...

...Protocols Streaking Bacteria on LB Agar Plate Over-Agar Antibiotic Plating You may also like... Making LB Agar...

...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...

...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...

... 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...

...amounts of liquid (think: 0.1 µL to 1 mL). When working in a laboratory, properly dispensing liquid ensures...plunger. Remove the entire pipette from the container making sure not to touch the sides of the container with...

...syringe and a blunt edge 18 ga Hamilton needle, taking care to avoid bubbles (Figure 1). 8 mL of 15% iodixanol...facing the 40% step. Collect up to 5 mL per tube, taking care to avoid collecting the proteinaceous material...

...Wear a lab coat, eye protection and gloves when working with this chemical. Note: If you add EtBr to your...the DNA. Note: When using UV light, protect your skin by wearing safety goggles or a face shield, gloves...

...the risk of contaminating reagents we recommend making small aliquots of master mixes, primers, and probes...wells. Cover the cartridge with the DG8 gasket, making sure that it is secure. Transfer the cartridge ...

... 30-37 ℃, 225 rpm for 1 hour. After the 1 hour shaking period, pool and gently mix the four tubes. Perform...transformation efficiency! Ensure that no arcing is taking place during electroporation. Arcing would manifest...

...wells. Cover the cartridge with the DG8 gasket, making sure that it is secure. Transfer the cartridge ...the risk of contaminating reagents we recommend making small aliquots of master mixes, and primers/probe...

...business of iPSC clone picking, a 100-fold decrease is the difference between picking 100 versus 10,000 clones...efficiency.   But what’s better: PAM-blocking or guide RNA blocking mutations? This depends entirely on...efficiency of guide RNA blocking mutations. So if the locus does not allow blocking the PAM sequence (e.g...intended mutation while taking advantage of the improved efficiency provided by blocking mutations. Essentially... incorporation of the blocking mutation. Despite the fact that both the blocking mutation and our intended... same CRISPR/Cas-blocking mutation but not the intended mutation. Here, the blocking-only templates will...used CRISPR/Cas9 extensively for the purpose of making sequence-specific changes in human induced pluripotent...