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...Postdoctoral Associations around Boston - From Panel Session 2: Defining the Postdoc: An Institutional Perspective...underrepresented groups under their mentorship with the same critical eye they place on others within their labs...

...module_attribute "schema_version" is_json="true" %}{% raw %}2{% endraw %}{% end_module_attribute %}{% module_attribute...seems obvious, it is important to remember that the same is true when designing gRNAs for using CRISPR technology...an alternative to HDR for introducing edits. The same locational constraints are even more exquisite for...concordance between different sequences targeting the same gene, suggesting that off-target effects did not...multiple gRNAs of different sequences give rise to the same phenotype in order to conclude that the phenotype...

...improvement in muscle physiological performance.  2. Trans-splicing Overview: For trans-plicing, splice...nbt0702-697 Hirsch ML, Agbandje-McKenna M, Jude Samulski R (2010) Little Vector, Big Gene Transduction:...Bellon I, Yin C, Chavala S, Pryadkina M, Richard I, Samulski RJ (2013) Oversized AAV Transductifon Is Mediated...

... iRFP713, % Oligomeric state Photo-stabilityc, t1/2, s pKa Brightness in HeLa cells vs. iRFP713, % d ..., and deep penetration of red-shifted light. The same NIR FPs and biosensors can be imaged at spatial ...

...variety of environmental conditions, and double every 2 hours. Yeast are also the first eukaryotic genome ... model organism in scientific research. Yes, the same kind we use in breads and other baked goods! Yeast...

..., whereas the mVenus-APT1 fusion does not (figure 2). In the mVenus-APT1, a lipidation motif, which is...conclusive evidence that the fusion operates in the same way as the native protein. Although a fluorescent...

... more from the authors in this blog post. Figure 2: iGluSnFR3 displays enhanced brightness, expression...to the range of concentrations expected in your samples. Higher affinity isn’t always better, especially...

...Nutr, 129 (2009a): 477–484. PubMed PMID: 10064313. 2. Chapman-Smith, A., & Cronan, J. E. “The enzymatic...pre-incubated with free avidin can be added to your sample. Any free biotin-binding sites on this complex ...

...15.4%) 46 (59%) of these students were in the last 2 years of their degree. Students filled in a pre-program... my mentor, since she does research at UK in the same field that I do: prostate cancer. She has guided...

...biology 14.6 (2004): 757-764. PubMed PMID: 15582400. 2. Tomkinson, Alan E., et al. "DNA ligases: structure...Watson-Crick base pair, are joined with nearly the same efficiency as a correct base pair. DNA ligases have... resistance-mediating polymorphisms in African samples of Plasmodium falciparum." Journal of clinical ...

...experience in cancer research After graduating with a 2:1 BSc in Molecular Biology (roughly a B average in...cancer and I loved the practical aspects of testing samples in the lab, but I really struggled with the large...

...effective recovery of knock-ins, but did result in 1-2 kilobases of additional sequence being introduced....changes 5’ to the PAM, sequences corresponding to the same strand as the PAM produced a higher editing efficiency...

... 10 90 2 x 10 8 10 of 2 x 10 8 dilution 90 2 x 10 7 10 of 2 x 10 7 dilution 90 2 x 10 6 10 of 2 x 10 6...6 dilution 90 2 x 10 5 10 of 2 x 10 5 dilution 90 2 x 10 4 10 of 2 x 10 4 dilution 90 2 x 10 3 Pro-Tip...molecules/μL To obtain a solution at 2 x 10 9 molecules/μL: 1.59 x 10 11 / 2 x 10 9 = 79.8X dilution ...your standard curve plasmid (2 x 10 9 stock made in step #1): Volume of 2 x 10 9 stock or previous dilution...stock 45 uL 10X 10X Dilution 2 5 uL Dil. 1 95 uL 20X 200X Dilution 3 20 uL Dil. 2 80 uL 5X 1000X Dilution ...empty NTC C AAV reference Sample 3 D E Sample 1 Sample 4 F E Sample 2 Sample 5 F Perform data analysis...virion). 5 μL sample + 39 μL H 2 O + 5 μL 10X DNase buffer + 1 μL DNase Gently mix sample (do not vortex...

...phosphate monobasic monohydrate (NaH 2 PO 4 ∙H 2 O), pH 7.0 138 g NaH 2 PO 4 ∙H 2 O 1 L deionized water Adjust...sodium phosphate dibasic (NaH 2 PO 4 ), pH 7.0 142 g of NaH 2 PO 4 ∙H 2 O 1 L deionized water Adjust pH...monobasic monohydrate (NaH 2 PO 4 ∙H 2 O) 610 mL of sterile sodium phosphate dibasic (NaH 2 PO 4 ) Store up to...purifying the same recombinant antibody. Section 2: Buffer exchange Choose Option 1 or Option 2 based on the...antibody sample. Pro-Tip If the volume of the sample is greater than 4 mL , then divide it into 2 columns...concentration of the pooled sample is below 1.0 mg/mL proceed to Option 2 with a buffer exchange/concentration...Centrifuge at 1000 x g for 2 min to collect the sample. Discard column after use. Determine antibody concentration...

...430825, 175 cm 2 Cellstack 5, Corning 3319, 3180 cm 2 Cellstack 2, Corning 3269, 1272 cm 2 Heat-inactivated...: 50 mM Tris HCl, 150 mM NaCl, 2 mM MgCl 2 Add the following to the 2 L sterile bottle: 1836 mL deionized... solution can be stored at 4 °C for up to 2 months. After 2 months, discard the tube and thaw a new working... mL of PBS. Aspirate PBS and add 2 mL of 0.05% Trypsin/EDTA. Wait ~2 min. Neutralize trypsin by adding...(CS5) (Link opens in a new window) (3,180 cm 2 - the same surface area as 21 x T-175 flasks). Cell stacks...flasks. Workflow Timeline Day 0: Seed cells in CS2 Day 2: Seed cells in CS5 Day 3 (am): Transfect cells Day...Biological Safety Cabinet 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel...

...Activation 95 10 2 1 Denaturation 94 0.5 2 40 Annealing/Extension 60 1 2 40 Enzyme Deactivation 98 10 2 1 Hold ...diploid cells, thus the reason for multiplying by 2. $$V = 2*{copies\ RRE \over copies\ RPP30}$$ Use the viruses...integrated copies of RRE. Since the samples that are assayed are diluted 2-fold serially, the concentration...factor of 2 across the dilutions. RPP30 copies should be relatively constant across samples. In the RRE...RRE example below, 2-fold serial dilutions of a sample were loaded in wells B01-H01. As shown in the image...Figure 2) below, the concentration of RPP30 positive droplets stays relatively even across samples (green...ddPCR Lentivirus sample data, RRE positive (blue) and negative (black) droplets Figure 2: ddPCR Lentivirus...

...materials D.2 Screening for inserts E. Producing Lentiviral Particles E.1 Recommended materials E.2 Protocol...Published articles H.2 Web resources I. Appendix I.1 Sequence of pLKO.1 TRC-Cloning Vector I.2 Recipes I.3 Warranty...VWR: #7177-48-2. Use at 100 μg/mL. Carbenicillin VWR: #80030-956. Use at 100 μg/mL. C.2 Annealing Oligos...oligo 5 μL Reverse oligo 5 μL 10x NEB buffer 2 35 μL ddH 2 O Incubate for 4 minutes at 95°C in a PCR machine...buffer 1 1 μL AgeI add ddH 2 O to bring to 50 μL final volume Incubate at 37°C for 2 hours. Purify with Qiaquick...buffer for EcoRI 1 μL EcoRI 14 μL ddH 2 O Incubate at 37°C for 2 hours. Run digested DNA on 0.8% low melting... For a standard T4 ligation, mix: 2 μL annealed oligo from step C.2 20 ng digested pLKO.1 TRC-cloning ...

... Example for AR0018 (lane 2 in Figure 1): Sample Peak 1 (contaminant) Peak 2 (contaminant) Peak 3 (HC)...follows: Figure 2 Using the box tool, draw a box around the entire first gel lane (as in Figure 2). Select Analyze...choose Use Equation . Select the Show R 2 checkbox. Pro-Tip The R 2 of the trendline should be between 0.95...of 4X sample buffer to each sample. Add 2 µL 10X reducing agent to each sample. Spin the sample briefly...Equipment Heat block 1–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel...bottom part of the gel where dye is visible. Section 2: Staining the Gel Place the gel in a plastic tray .... Figure 1 Recombinant antibody preps should have 2 clear bands at ~50 kDa and ~25 kDa corresponding to...

... 95 10 2 1 Denaturation 95 0.5 2 50 Annealing/Extension 60 1 2 50 Signal Stabilization 98 10 2 1 Hold ...factor of 2 across the dilutions. In the example below, 2-fold serial dilutions of a sample were loaded...AAV sample, and the three final dilutions are assayed. The samples that are assayed are diluted 2-fold... considered biosafety level 1 but may require BSL-2 handling depending on the insert. Please ensure that...single channel pipette 1–10 µL multichannel pipette 2–50 µL multichannel pipette 20–200 µL multichannel ...20X): 5 µL in 95 µL 1X PCR buffer (1:20) Dilution 2 (20X): 5 µL in 95 µL 1X PCR buffer (1:400) Dilution... DG8 cartridge into the cartridge holder. Using a 2–50 µL multichannel pipet, load 20 µL of the reaction...

...PCR tube Ice Bucket 2 μL Template DNA (10 ng-500 ng) 5 μl 10X Taq buffer with MgCl 2 1 μl dNTP mix (10 ... reagents on ice): 2 μL Template DNA (10 ng-500 ng) 5 μl 10X Taq buffer with MgCl 2 1 μl dNTP mix (10 ...normally sterile dH 2 O. To make a 100uM stock of any primer, add a number of µl of dH 2 O equal to the number...annealing temperature step-wise by 1-2°C. The rate of DNA synthesis is ~1-2 kb/min. The extension time can ...work adequately. Divalent cations such as Mg 2+ and Mn 2+ stabilize the buffer solution. These cations...(PCR). Basic PCR Program Initial Denaturation for 2 minutes at 94°C: This initiation step heats the double...target DNA strand accurately and rapidly. Repeat steps 2-4 25-30 times. Final Extension for 5 minutes at 72...