How to Test Positive Results for Phage Induced Nutrient Agar Plates
- Learn how to test positive results for phage-induced nutrient agar plates
- Step-by-step instructions for detecting immunopositive signals
- Use affinity-purified anti-calcineurin antibody and alkaline phosphatase-conjugated rabbit anti-goat IgG for detection
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先ほどの続きです。
悩んでいる 個所は以下のところです。 手順が想像つかないもので、わかる方よろしくお願いいたします。 nutrient agar plates,seeded with 20000 plaque-forming units of phage were induced and then overlaid with IPTG-saturated nitrocellulose filters. duplicate lift of 3hr and 5hr were washed in 50mM Tris-Hcl containing 150mM Nacl(tris-burrered saline;TBS)and then incubated tor 20 min with blocking solution(5%nonfat dry milk in TBS plus 10mM soduim azide and 0.01%thimerosal). affinity-purified anti-calcineurin antibody that had been adsorbed with immobilized E.coil lysate was diluted into blocking solution and incubated with the plaque-containing filters for 2.5hr. after washing with TBS plus 0.05%Tween20,filters were incubated for 1hr with alkaline phosphatase-conjugated rabbit anti-goat IgG and washed as described above. immunopositive signals were detected with the chromogens nitroblue tetrazolium and bromochloroin-dolyl phosphate,and the corresponding plaques were excisedwith sterile pipette tips. after overnight elution of phage in 1mM of 10mM tris-hcl plus 10mM Mgcl2.a portion was diluted 1:2000and 20μl was used to seed nutrient plates for testion of positives.
- hatimitsuboy
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- 生物学
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前回も回答したので再登場です（笑）。 論文の一部でしょうか？ 抗体を用いたライブラリーからのスクリーニングですね。 この方法でしたら、分子生物学の基本ですので、 多くの実験書や教科書に図入りで記載されていると思います。 ただし、最新の方法を中心とした本には載っていないかもしれません。 簡単にいうと、発現ファージベクターを用い、 lacZとの融合タンパク質としてcDNAを発現させ、 抗体によりスクリーニングする方法です。 Molecular Cloning (A Laboratory Manual) という実験書があるのですが、 かなり詳細に説明されています。理系の大学でしたら、図書館や研究室には 必ずあると思いますよ。特に「Screening Expression Libraries with Antibodies and Oligonucleotides」の章を読んでみて下さい。
関連するQ&A
- 和訳 さきほどの続きですが...
さっきものすごい長い和訳をお願いしたのですが、１つでも余りにも長すぎたので２つに分けました！！ この和訳もお願いします(>_<) Where should you start to learn about keeping fish? Check out a book or two from your library or surg the Internet. You will need to know about things like filters, water quality and the types of tanks available that best suit your needs. Also, you must decide whether you want to have saltwater or freshwater fish. Spend some time in pet shops and hobby shops to see what is available. Generally, a beginner should start with a freshwater aquarium. Salwater fish can be pleasant and interesting to look at, but the water chemistry is much more complex. In addition, saltwater fish are difficult to raise and are often collected in the wild, sometimes in environmentally unfriendly ways.
- ベストアンサー
- 英語
- 生物英文質問
This was determined by adding a modified Bray's powder to an aliquot placed in a spot plate. This powder was made by eliminating MnSO4 and zinc dust from the Bray's nitrite-nitrate powder. 「スポットプレートに改良したBrayの粉を加えた」 とありますが、スポットプレートとはどういうもので Brayの粉とはどんな物なのでしょうか。 A series of diluted enrichment cultures was plated onto the acetamide medium solidified with 1.5% Noble agar. 「Noble(人名？)の寒天で固めたアセトアミド培地 の上に集積培養をした菌を塗った」 ・・・ですか？ Individual colonies were subcultured on acetamide agar plates for 5 days and tested for nitrite production by adding nitrite reagent. 「各コロニーはアセトアミド寒天培地で ５日間継代培養され、亜硝酸試薬を加え、 亜硝酸性生物の試験を行った」 という感じでしょうか？ These colonies were streaked again on acetamide agar and nutrient agar plates. 「これらのコロニーは、アセトアミド寒天および 寒天培地プレート上で再び兆候が見られた。」 ここに来て、培地がに２種類に増えたような気がするのですがどうなのでしょう・・・？
- ベストアンサー
- 生物学
- Ｍａｔｅｒｉａｌｓ・Ｃｕｌｔｕｒｅｓの訳
物質と培養の方法についての部分です。 全訳ではなく、まとめるための訳です。 間違いや、不足分のご指摘をお願いします。 Materials.------------------------- PO was synthesized by the method of Quastel et al. and sterilized with 0.20-μm (pore size) membrane filters. Hydroxylamine hydrochloride was recrystallized from ethanol-water. Solutions of hydroxylamine were prepared and neutralized immediately before use. Enriched media and agar were from Difco Laboratories, Detroit, Mich. Cultures.------------------------------ The bacteria studied are listed in Table 1. Cultures were maintained on agar plates containing the tryptic soy-yeast extract medium (minus nitrite and nitrate) of Grant and Payne. Cultures were transferred from these plants to agar plates containing 7 mM PO-0.05% yeast extract-mineral salts (PO-YE) medium. 素材－－－ POはQuastelらの方法を用い、0.20μmの薄膜フィルタによって殺菌されました。 塩酸ヒドロキシルアミンはエタノール－水系から再結晶しました。 ヒドロキシルアミンの溶液は使用直前に準備し、中和しました。 豊かになった媒体と寒天はDifcoの実験室からでした。 （Difcoは人名でしょうか） 培養ーーー 研究した細菌は、表１に示します。 菌は、トリプシンの大豆酵母エキスを含む寒天プレートに保持されています。 （soyは大豆？） 菌をプラントから７mM PO 0.05％ 酵母エキス無機塩類培地を含んでいる寒天プレートに移しました。
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- 化学
- 英語論文の訳
まとめるのが目的なので、全訳はしていません。 「大事なのに抜けている」など補足をお願いします。 The mineral salts mixture was that of Verstraete and Alexander (24). Individual colonies on PO-YE plates were used to inoculate liquid PO-YE medium or a liquid medium consisting of 15 mM PO-mineral salts (PO-MS) medium. Liquid cultures (100 ml) were incubated in 500-ml Erlenmeyer flasks with a gyratory shaker rate of 280 rpm to assure adequate oxygenation (8). Secondary liquid cultures used for quantitative measurements were inoculated by a 100-fold dilution of a previous culture which was in late exponential growth. Cultures were incubated at 30℃ except for Neiserria sicca, which was incubated at 37℃. Media used for agar plates were solidified with 1.8% agar. PO-YEプレートのコロニーは15 mM PO無機塩類培地からなる液状のPO-YE培地か液体培地に摂取するために使用しました。 液状の培地は適正な酵素処理をするために、280rpmの旋回シェーカーの500ml三角フラスコで作られました。 培養は30℃でNeiserria siccaを除いて行われ、37℃で増殖させました。 18%の寒天で凝固しました。
- ベストアンサー
- 生物学
- 先程の続きです。 日本語訳お願いします。
Einsten often said that his most famous theory was also inspired by music, its forms and relationships. It seems that certain physical actions stimulate the brain,, create connections, and speed up thought. We have all had the experience of being literally unable to sit still when we are trying to resolve something in our minds. We pace up and down or shake a leg, almost as if such nvoluntary motions were needed to move our thoughts forward. Sometimes the mind-body relationship works a bit differently: while taking a long walk or rowing a boat across the lake, just letting our thoughts. drift, we suddenly receive from the subconscious, without effort, the solution to a problem that had been troubling us for weks. I personally know that playing the piano has this effect; maybe it has something to do with both sides of the brain being stimulated by the independent movements of both hands. In any case, I have to keep a notebook handy while practicing, because I tend to get the answers to all kinds of questions ranging from the insignificant (where I left the car keys) to the significant (my life goal for the next decade). This mind-body unity is only one of the rewards to be found in making music, at any level, as an adult. Ironically, far too many adults who study music under the general category of "amateurs," are so modest in their expectations of mastering an instrument that these rewards turn .out to be much less than what they might have been. The paradox is that adult music learners, while they often have the lowest expectations, are in a uniquely excellent position to succeed. Many take up an instrument with warmhearted desire, love of music, leisure time and extra money to support the lessons. Even more importantly, they approach music with more maturity and intelligence. But how much can adults learn if they are constantly comparing themselves negatively to others (children, no less) and feeling guilty about every unmusical wrong note? Add to this the mistaken notion of practice as boring discipline, acceptance of the traditional "no-mistakes" approach to learning, and uncertainty that they have any real talent for music, and they soon find themselves in a state of semi-paralysis of body and mind. Even the word amateur has conflicts of meaning. While it literally means "lover" in French, it can also carry the connotation of dabbler, a person who is somehow fated never to be very good. To describe someone's work as amateurish usually is not a compliment. But amateur status can be joyous-embracing free choice, pure love of what one is doing, and endless possibilities for discovery. (Adapted from a work by William Westney)
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- 英語
- 英文の訳
These were prepared by electric-spark machining, with a cross section of 5×5mm^2 and a gauge length of 23mm for the working part, and a cross section of 5×10mm^2 and a length of 10mm for each of the two button heads. 「これらの試料は放電加工機によって準備され、～～～～。」 この文ですが、試料の形状を説明しているのですがどのような形になっているのかまったくわかりません。 cross section 断面積 gause length ゲージ長 だいたいでもいいのでこの文が訳せる方がいましたらお願いします。
- 締切済み
- 英語
- 和訳をお願いします。
The forts were triangular or quadrangular and built of concrete, with a surrounding ditch and barbed-wire entanglements. The superstructures were buried and only mounds of concrete or masonry and soil were visible. The forts were armed with 78 x 210-millimetre (8.3 in) howitzers, 150-millimetre (5.9 in) and 120-millimetre (4.7 in) guns and 57-millimetre (2.2 in) quick-firers. Each fort had retractable cupolas, mounting guns up to 6-inch (150 mm) and the main guns were mounted in steel turrets with 360° traverse but only the 57-millimetre (2.2 in) turret could be elevated. The forts contained magazines for the storage of ammunition, crew quarters for up to 500 men and electric generators for lighting. Provision had been made for the daily needs of the fortress troops but the latrines, showers, kitchens and the morgue had been built in the counterscarp, which could become untenable if fumes from exploding shells, collected in the living quarters and support areas as the forts were ventilated naturally. Pentagonal Brialmont fort, 1914 The forts were not linked and could only communicate by telephone and telegraph, the wires for which were not buried. Smaller fortifications and trench lines in the gaps between the forts, to link and protect them had been planned by Brialmont but had not been built by 1914.
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- 英語
- her eyes were a shuttered, washed-out blue」というところと、２つめの文全体がわからないです。日本語を教えてください。
英語の２つの文の意味がよくわからずにおります。「her eyes were a shuttered, washed-out blue」というところと、２つめの文全体がわからないです。日本語を教えてください。英文は「In the other woman's blank face, her eyes were a shuttered, washed-out blue. The Universe watched as the painful moment played itself out, watched with a grim and inexorable regard that I was glad was fixed on A and the stranger rather than on me.」です。よろしくお願いいたします。
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- 英語
- 英文翻訳をお願いいたします。
The Meuse valley was a route by which France or Germany could be invaded and after the Franco-Prussian War, General Henri Alexis Brialmont fortified the valley at Liège and Namur, to deter France and Germany from violating Belgian sovereignty. The Fortified Position of Namur (FPN) was built from 1888–1892, about 7 kilometres (4.3 mi) from the centre of Namur, to a standard design of triangular and quadrilateral shapes, to minimize the number of defensive batteries in the fort ditches, with the point facing outwards. On the left bank of the Meuse lay the modernised forts of Fort de Malonne, Fort de Saint-Héribert and Fort de Suarlée, the unmodernised Fort d'Emines and Fort de Cognelée and the modernized Fort de Marchovelette. On the right bank were Fort de Maizeret, Fort d'Andoy and Fort de Dave, all modernized. The obsolete Citadel of Namur in the town became redundant. The forts were built of non-reinforced concrete but this could only be poured in daylight, which caused weak joints between each pour. A citadel was built and covered by 3–4 metres (9.8–13.1 ft) of concrete; caserne walls which were less vulnerable, had concrete of 1.5 metres (4.9 ft) thickness, inside a defended ditch 8 metres (26 ft) wide. The entrance had a long access ramp at the rear facing Namur, protected by a tambour with gun embrasures perpendicular to the entry, a rolling drawbridge retracting laterally over a 3.5 metres (11 ft) pit equipped with grenade launchers, an entrance grille and a 57-millimetre (2.2 in) gun firing along the axis of the gate. The forts at Liège and Namur had 171 heavy guns, with each fort equipped with 5–8 Krupp guns of 120-millimetre (4.7 in), 150-millimetre (5.9 in) and 210-millimetre (8.3 in) calibre, which were the most modern armaments available in 1888, mounted in retractable armoured steel turrets made in France, Belgium and Germany. Three smaller retractable turrets were built in the triangular forts and four in the quadrilateral forts, with 57-millimetre (2.2 in) guns for short-range defence and 6–9 more 57-millimetre (2.2 in) guns were mounted in casemates to defend the ditches.
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- 英語
- 主語はどれでしょう？
下の文を読んでいるのですが、'to which has been added a discrete nutrient'の部分で、has been addedの主語になっているのは、どの語句でしょう？ a discrete nutrientではないかと思うのですが、意味は通るもののなぜ動詞のあとになっているのかわかりません。別の読み方があるのでしょうか。 (f) A product within a class of food for which a nutritional quality guideline has been established and to which has been added a discrete nutrient either for which no minimum nutrient level or nutrient range or other allowance has been established as appropriate in the nutritional quality guideline, or at a level that exceeds any maximum established as appropriate in the guideline, shall be ineligible to bear the guideline statement provided for in paragraph (b) of this section, and such a product shall also be deemed to be misbranded under the act unless the label and all labeling bear the following prominent and conspicuous statement: ‘‘The addition of lll to (or ‘‘The addition of lll at the level contained in) this product has been determined by the U.S. Government to be unnecessary and inappropriate and does not increase the dietary value of the food,’’ the blank to be filled in with the common or usual name of the nutrient( s) involved. http://edocket.access.gpo.gov/cfr_2002/aprqtr/pdf/21cfr104.5.pdf
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- 英語
お礼
前回に引き続きありがとうございました。 参考書まで紹介してくださってありがとうございます。 早速図書館で確認してみたいと思います。