अब विधायक राणा रोज करेंगे कोरोना मरीजों को वीडियो कॉल, जानेंगे हाल
आरसीएफ में एलपीजी व कार्बन डाईऑक्साइड गैस प्लांट का सेफ्टी ऑडिट
कार-ट्रक की टक्कर में कार सवार दो लोगों की मौत
कार और बाइक की भिड़ंत में महिला की मौत, पति जख्मी
फिरोजपुर में एक रेस्टोरेंट में पिलाई जा रही शराब, वीडियो वायरल
काला संघिया गोलीकांड : मुख्य आरोपी समेत दो गिरफ्तार
तेज आंधी से चिमनी और सोलर पैनल दूर जाकर गिरे
बाजार खुले लेकिन ग्राहक गायब, मजबूरी में ऑटो वाले बेच रहे सब्जी
कपूरथला की माडर्न जेल से 495 कैदी जमानत-पैरोल पर रिहा
जालंधर से तीन ट्रेन में आजमगढ़ व बिहार के श्रमिक रवाना
आरएमपी डाक्टर को धमकाकर 30 हजार वसूले, दो पत्रकार गिरफ्तार
पंजाब में कोरोना महामारी से बुधवार को दो और लोगों ने दम तोड़ दिया। इसके साथ ही प्रदेश में संक्रमण से मरने वालों की संख्या 27 हो गई।
जालंधर से लखनऊ के लिए श्रमिक ट्रेन 14 मिनट की देरी से बुधवार को सुबह 11:14 बजे रवाना हो गई।
पंजाब सरकार ने कोरोना वायरस से निपटने के लिए केंद्र से 150 करोड़ की मांग की थी।
राशन किट के लिए एसडीएम दफ्तर में उमड़े सैकडों लोग
कोटकपूरा में बारिश और ओलावृष्टि, भीगा गेहूं
मोगा में कोरोना वायरस की चेन लगातार बढ़ती ही जा रही है।
थाना सात के क्षेत्र मिठ्ठापुर में तेज रफ्तार ट्रक की चपेट में आने से एक्टिवा सवार युवक की मौत हो गई।
सिविल अस्पताल में मेडिकल कराने उमड़े प्रवासी, कर्फ्यू की उड़ीं धज्जियां, लोगों के सामने हाथ जोड़ते नजर आए विधायक
पंजाब में जून में धान की रोपाई शुरू होने के आसार हैं, लेकिन इस बार धान की रोपाई के लिए श्रमिक कहां से आएंगे।
पंजाब से करीब साढ़े नौ लाख श्रमिक अपने गृह जिलों में रवाना होने के लिए लगभग तैयार हैं।
पंजाब में पेट्रोल और डीजल बुधवार सुबह छह बजे से दो रुपये महंगा हो जाएगा।
मुख्यमंत्री ने यह भी स्पष्ट किया कि वे संबंधित देशों द्वारा पंजाब लौट रहे लोगों को दिए जाने वाले मेडिकल प्रमाणपत्रों पर यकीन नहीं करते।
भूजल की बचत और कर्फ्यू/लॉकडाउन की वजह से किसानों को मजदूरों की कमी से निपटने में भी मदद मिलेगी।
ट्रेन को जालंधर सिटी रेलवे स्टेशन से डीसी वरिंदर कुमार शर्मा और पुलिस कमिश्नर गुरप्रीत सिंह भुल्लर की निगरानी में रवाना किया गया।
होमगार्ड की पत्नी-बेटों ने डिपो होल्डर के भाई को पीट-पीटकर मार डाला
This protocol describes a simple single-step column purification (SSCP) of rAAV2 by gravity flow based on its affinity to heparin, without ultracentrifugation.
This protocol is used to determine the concentration of DNase-resistant vector genomes (i.e., packaged in the capsid) in purified recombinant adeno-associated virus (rAAV) preparations. The protocol begins with treatment of the vector stock with DNase I to eliminate unencapsidated AAV DNA or contaminating plasmid DNA. This is followed by a heat treatment to heat-inactivate DNase I, to disrupt the viral capsid, and to release the packaged vector genomes for quantification by real-time polymerase chain reaction (PCR) using a set of standards (linearized plasmid used for vector production) containing known copy numbers. To accomplish high-throughput titration, the primer and probe sets used in real-time PCR are usually designed to target common elements present in most rAAV genomes, such as promoters and poly(A) signals. This strategy significantly reduces the number of PCRs, controls, and turnaround time. Several important controls should be included in the assay as follows: The first two controls should have a known copy number of the rAAV genome plasmid treated or not treated with DNase I. This control tests the effectiveness of DNase treatment. To control for potential cross-contamination between samples during the preparation process, a blank control containing nuclease-free water only should be processed and tested in parallel. A validation vector sample with a known titer should be included in every assay to monitor interassay variability. Finally, for the PCR run, a no-template control (NTC) is included to indicate cross-contamination during PCR setup.
Adeno-associated virus (AAV) recombinants are currently the vector of choice for many gene therapy applications. As experimental therapies progress to clinical trials, the need to characterize recombinant adeno-associated viruses (rAAVs) accurately and reproducibly increases. Accurate determination of rAAV infectious titer is important for determining the activity of each vector lot and for ensuring lot-to-lot consistency. The following protocol developed in our laboratory uses a 96-well TCID50 format and quantitative polymerase chain reaction (qPCR) detection for the determination of rAAV infectious titer.
High-throughput sequencing methods have created exciting opportunities to explore the regulatory landscape of the entire genome. Here we introduce methods to characterize the genomic locations of bound proteins, open chromatin, and sites of DNA–DNA contact in Xenopus embryos. These methods include chromatin immunoprecipitation followed by sequencing (ChIP-seq), a combination of DNase I digestion and sequencing (DNase-seq), the assay for transposase-accessible chromatin and sequencing (ATAC-seq), and the use of proximity-based DNA ligation followed by sequencing (Hi-C).
β-Galactosidase has been used extensively both as a label in enzyme immunoassays and for immunocytochemistry. One good substrate is 5-bromo-4-chloro-3-indolyl-β-
The repetitive immunization at multiple sites (RIMMS) protocol capitalizes on the animal's innate immune system, which is genetically preprogrammed to recognize many antigens. By repetitively immunizing the animal, B cells that recognize the antigen are kept continuously expanding until the lymph nodes are harvested for hybridoma generation. This is a good method for making a more diverse repertoire of antibodies or antibodies directed against conformational epitopes.
Subtractive immunization is useful for directing the immune response away from dominant epitopes to allow antibodies against less immunogenic antigens to be developed. It is also useful when there is a need to downplay the response to carriers added to peptides and recombinant proteins, such as Gst and Ig, to aid in production and purification. Subtractive immunization protocols also can be helpful if the target protein is part of a mixture (such as a cell lysate) or present on the surface of transfected cells.
The Klenow fragment, which retains the template-dependent deoxynucleotide polymerizing activity and the 3' -> 5' exonuclease of the holo-enzyme but lacks its powerful 5' -> 3' exonuclease activity, is used to fill recessed 3' termini of dsDNA. In this protocol, fragments suitable as templates for the end-filling reaction are produced by digestion of DNA with an appropriate restriction enzyme. The Klenow enzyme is then used to catalyze the attachment of dNTPs to the recessed 3'-hydroxyl groups.
Escherichia coli DNA Pol I can carry out three enzymatic reactions: It possesses 5' -> 3' DNA polymerase activity and 3' -> 5' and 5' -> 3' exonuclease activity. Pol I can be cleaved by mild treatment with subtilisin into two fragments; the larger fragment is known as the Klenow fragment. The Klenow fragment retains the polymerizing activity and the 3' -> 5' exonuclease of the holo-enzyme but lacks its powerful 5' -> 3' exonuclease activity. These enzymes and their applications in molecular cloning are introduced here.
Stocks on Wall Street jumped Friday despite historic job losses suffered by the economy. Fred Katayama reports.
Piper Sandler senior technical analyst Craig Johnson says the S&P 500 may trade in a narrow range through the summer before breaking north.
The steep plunge in U.S. payrolls caused by the pandemic also slammed the healthcare sector, as many workers in dentists' and doctors' offices lost their jobs. Fred Katayama reports.
While U.S. government guidelines say jobless workers who qualify for assistance should get payments within three weeks of applying, many - like Florida resident Claudia Alejandra - have been waiting twice that long. This report produced by Yahaira Jacquez.
Neiman Marcus Group filed for bankruptcy protection on Thursday, marking one of the highest-profile collapses yet among retailers forced to temporarily close stores in response to the COVID-19 pandemic. Freddie Joyner has more.
Wall Street's indexes climbed Thursday, with the Nasdaq erasing losses for 2020, following a clutch of upbeat earnings reports led by PayPal. Fred Katayama reports.
H&M, the world's second-biggest fashion retailer, said local currency sales have tumbled 57% since the start of March compared with a year ago. Ciara Lee reports
Payne Capital Management's Courtney Dominguez says the high level of investor pessimism indicates the markets may have bottomed. She says savings could make their way into stocks.
U.S. cable firms are in for pain even after the pandemic starts to fade. Sports rights to air football games and other matchups are expected to soar and so will monthly bills. Jennifer Saba explains why non-sports lovers will choose Netflix and Disney+ and ditch their cable.